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thank you for joining the Jackson

laboratories presentation on protector

research no your be six miles my name is

Peter Kelman s'en and I've been with

technical information services at the

Jackson laboratory for over 11 years c57

black 6 or b 6 inbred mice the most

commonly used mice in biomedical

research this inbred strain is the most

well characterized the first to have its

entire genome sequenced and is the

genetic background strain of choice for

most targeted mutations and transgenics

the universal acceptance and demand for

B 6 mice has necessitated their

production from multiple sources

introducing genetic and phenotypic

variability that has very important

consequences for accurately interpreting

and repeating research results so if we

just did a quick exercise and just

looking at these three mice Mouse trains

if I were to ask you which of these two

are genetically the most similar a and B

B and C or a and C most of you would

likely answer B and C as they both look

like c57 black 6 mice they're both black

however it turns out that actually a and

B are the most genetically similar the

strain a is a mutation that arose in the

colony of c57 black 6 J that makes them

I cell by now but it's a single gene

mutation while as the c57 black 6 J

which is a picture B and the c57 black 6

and J my switches picture see those two

strains have been separated since 1951

and they differ in many different

mutations that are involved in

metabolism neural biology immunology

vision hearing as well as behavior and

that's really the heart of what I want

to talk to you about today is is really

knowing what black 6 sub strain of mice

you were using so that you can best

interpret your data just as another

example to drive that home in that coat

color is not actually a very good

indicator of genetic background these

are almost strains that are all c57

black 6 J backgrounds and not only do we

have the albino ones but we have revert

ins to a GU T beige the dominant

spotting phenotype so all five of these

strains are all c57 black state

backgrounds but they all have a

different coat color so again coat color

is not a very good indicator of genetic back

back

ground really you would actually have to

survey the the actual genome really to

determine which strains are most closely

related the Jackson Laboratory we do

have two substrates of c57 black 6 we

have both the c57 black 6 J which is our

stock is zero zero zero six sixty four

and the c57 black 6 NJ which is our

stock is zero zero five three zero four

both of those colonies are maintained at

a high health status they're very well

characterized strains some of the most

published strains that you can find and

we have extensive phenotypic data

available as well as consistent data

reproducibility which I will talk to you

a little bit later when I get into it

discussing our GSP program before we

start to talk about different sub

strains of c57 black 6 we really need to

look at the origin of the c57 black 6

inbred strain itself to help understand

the genetics of that before we can talk

about the genetics of substrates so back

in the early part of the 1900s is and

earlier there was a lot of people who

sort of raised mice they were called

most fanciers they raised mice they had

different interesting coat colors or

different phenotypes and and they would

trade them amongst each other or sell

them and one of them her name was Abbie

Lathrop and she was in Massachusetts and

she had she provided mice to the Bussey

Institute at Harvard University and was

also about that time that geneticists

really realized that mice were an ideal

model organism for studying mammalian

genetics I mean they're small they're

easy to maintain they generally have

really good reproductive performance and

their anatomy and physiology was very

similar to that of humans and then again

really at the beginning of the 20th

century dr. William castle started using

mice from Abbie Lathrop as well as a lot

of other people and he again was at the

bus the Institute in Harvard and then

shortly they're late after that one of

his students he see little who was the

founder of the Jackson Laboratory he

began in breeding those mouse stocks as

a student of dr. castle and really of

course that is the initial inbreeding

which led to a lot of the classical

inbred strains that we have today and in

1921 tsetse little got some of mammy

lathers pet shop stock in this case it

was a mouse train that has a waltzer

phenotype it's a mutation that sort of

the spinning dancing behavior and when

he made it female number 57 two

different males he ended up getting

black mice and the other he got brown

mice and as he started to inbred there

was a mutation arose in the brown colony

that led to a legend or gray phenotype

and so as those mice were inbred they

were called c57 because was female

number 57 and then BL for black BR for

brown and L for lead as simple as that

so the way inbred mouse strains are

generated is by brother-sister mating

every generation so the progeny of your

initial cross which are the f1 hybrids

that generation you take a brother and

sister you breed them together to

produce f2 mice again you take a brother

or sister and you make them together to

produce f3 mice and then you would keep

on doing that and after 20 generations

you would generate an inbred mouse

strain and with inbred mice all the

individuals are virtually genetically

identical so you have a high degree of

genetic homogeneity and the mice are

homozygous at virtually every locus and

that way with that high degree of

homogeneity you're gonna have a high

degree of statistical reproducibility so

if you take a group of inbred mice and

you give them one treatment and you take

another group of the same inbred strain

and you give them a different treatment

or no treatment any of the differences

you see are going to be due to the

treatment not due to underlying genetic

differences as well as if you then

introduce mutations or transgenes

the mice that have the mutation are

transient as compared to mice that don't

because they're otherwise genetically

identical any differences you see will

be due to the presence of the

transgender that or the mutation not

again underlying genetic differences

that makes inbred mouse strains a very

very powerful tool for studying genetics

when you are working with inbred mouse

strains there are really two things that

you need to make sure you're very well

aware of you need to understand the

basic genetics and mutations and

phenotypes of the inbred strain in

general and then as you see coming up

you also need to understand what sub

strain of that inbred strain you are

using and what mutations and phenotypes

might be specific to that sub strain

that way you can best interpret your

results correctly I'll give you an example

example

all c57 black 6 sub strains carry a

mutation in a

called kid here in 23 known as the

age-related hearing loss or AHL mutation

therefore all c57 black 6 mike says they

get older they will start to lose some

of their hearing and this of course is

going to cause a complication

interpreting data that you have that

influences diseases or phenotypes that

involve hearing and neurobiology

for example phenotypic analysis of genes

that are implicated in cognitive

behavior like fear conditioning and

older mice will require an auditory cue

which could be problematic than using

c57 black 6 mice and this mutation can

infect lots of researchers including

autism anxiety and stress disorders

addiction and cardiovascular functions

so it's really critical that you

understand the basic mutations and

phenotypes of the inbred strains you are

using but then you also need to look at

the sub strains and realize that sub

strains can develop relatively quickly

and so one has a sub strain when a

colony is separated by 20 or more

generations or phenotypic or genetic

differences are discovered and it's

relatively quick to generate these sub

strains so if you have a c57 black 6

parental colony and lab a gets those

mice and breeds them for 10 generations

and lab B gets the same mice and breeds

them for 20 generations those two

generations add up and the mice held by

lab and lab B are actually 30

generations apart so it can actually go

a lot quicker than you expect and see

see little who founded the Jackson

Laboratory and also the one who created

the c57 black 6 inbred strain and since

he was at the Jackson laboratory the c57

black 6 J then is the parental colony

but between 1930 and 1970 see see little

sent many of those c57 black mice to

researchers and institutions around the

world because he truly believed that an

important part of research is sharing

resources to make that research more

robust and so he certainly did that and

then you can see that many different c57

black 6 sub strains have been

established around the world and in 1951

some of the black 6j mice were sent to

nih and they became the black 6n mice

those are the two sub strains I'm going

to talk

a lot of it about today because those

are probably the most two commonly used

sub strains of c57 black 6 that are used

in biomedical research so in addition to

the c57 black 6 j sub strain and the c57

black 6 and sub strain the jackson

laboratory actually has additional sub

strains that have been in various places

and and what you'll find is everything

after the 6 is going to be the substrate

designation and that's going to use

laboratory codes the J is the laboratory

code for the jackson laboratory and is

the laboratory for the nih and then you

can see that for example we also have

the c57 black 6 h aj which went to a dr.

Hauschka and then came to the jackson

laboratory the byj had gone to dr.

bailey and then the jackson laboratory

and then the e IJ went to dr. Iker and

then back to the jackson laboratory and

then of course the j is at the end of

all those because we are the lab that is

currently maintaining the strain and if

you are interested in tracing laboratory

codes there is this link at the bottom

of the page to the Institute for

laboratory animal research or I'll are

you can look up any lab code or request

a lab coat of your own if you do need

those and I've mentioned a couple times

now already that not all c57 black 6

substrates are the same and that they

will differ genetically in a variety of

ways whether it be single nucleotide

polymorphisms or snips there can be

insertions deletions also known as in

Dells there could be copy number

variation either due to duplications of

regions of the chromosomes and all of

these would be some more and more

spontaneous mutations and these genetic

differences can translate into

phenotypic differences between different

c57 black 6 sub strains they can affect

a variety of phenotypes including those

in metabolism and I'll give you some

examples from neurobiology immunology

and others as well so here's a an

example of comparing how the c57 black 6

j and the c57 black 6 NJ mice respond to

a high respond to a high-fat diet also

known as diet induced obesity or do in

this case the mice were fed a 60 kcal

percent high-fat diet

starting at 6 weeks of age and as you

can see on the graph at the right this

c57 black 6 j mice gain more weight than

the c57 black 6 and

but I'd like you to imagine a scenario

well let's say you had a mutation or

trans gene on a c57 black 6 background

and you put them on a high-fat diet and

what if your data sort of came in

between those two graphs well if you

were comparing your data to that for the

black 6j if that's what you were using

as controls that would indicate that

your mutation or tran gene had reduced

the amount of fat put on but if you were

using c57 black 6 and jameise you may

then believe that your mutation or transient

transient

increase the amount of fat so depending

on the control that you use the way in

which you interpret your data will

differ and that's why it's really

critical to understand not only what

stream you're using but what substrain

of c57 black 6 are you are using so you

can properly interpret your data and

share that with the world here's again

another example from the same reference

comparing a glucose tolerance test

between c57 black 6 j and the black 6 NJ

also on a high fat diet and glucose

tolerance test really measures the

ability of the mice to clear glucose

from the blood so they're given a bolus

of glucose and then you measure the

blood glucose levels over time and what

you can see is that the black 6j mice

even a couple weeks as well as it gets

more prowess after 14 weeks on the

high-fat diet can't clear glucose as

well as the black 6nj mice and again the

same situation if your data fell in

between those depending on what you

chose as your controls would determine

how you interpret the data and if you

choose the wrong sub strain of black 6

you may incorrectly interpret that data

and one can also see neurological

differences as well when one looks at

mice from the C 50 some black 6j mice

compared to the black 6n from Charles

River those two strains are wild-type

for a gene called SN CA or it's a alpha

synuclein protein but when you look at

the black six months from harlan those

mice have had a spontaneous deletion of

that gene doesn't necessarily give a

visible phenotype but the alpha

synuclein protein is implicated in a

wide range of neurodegenerative diseases

and it's that primary structural

component of the Lewy bodies that are

found in the brains of Parkinson's

disease patients so that mutation

doesn't necessarily give a obvious

phenotype however if you are studying

some sort of neurodegenerative disease

that may affect the data depending on

your experiment on using which substrate

you've use so it's really important

again to understand not just a strain

you're using but the substring of c57

black 6 so that you can either choose

the right substrate for your experiments

or at least interpret your data

correctly and here's another example of

a behavior in which the black 6 mice

which are known to have a preference for

alcohol over other strains so if you

give them a choice of water with alcohol

or water without they'll prefer the

water with alcohol but you can see that

the black 6j mice actually will drink more

so they'll consume more alcohol and

they'll have a higher preference for it

than the black 6n mice which again leads

to a different phenotype again with two

different sub strains again depending on

what controls you use in your experiment

you may interpret your data can

correctly depending on your strain but

also there's another example of having

two different sub strains which have a

different phenotype which has some

notable differences now in gene

expression as you can see between the

genes at the graph in the lower right

hand corner of plaque 9 and the d40 me

RTD 44 4 9 II which could potentially be

responsible for those different

phenotypes so there's different sub

strains not only is an issue to pay

attention to but can be potentially good

experimental models for looking at those

different phenotypes and determining the

underlying genetic differences and the

last neurological one involves vision in

this case it's a gene known as CR b1

which is local localized to mulher cells

and the photoreceptor inner segments

mutations in that gene is associated

with retinal diseases in people

including retinitis pigmentosa as well

as retinal degeneration there's a

mutation in that gene called RT 8 for

retinal degeneration 8 that results in

this progressive spotty retinal

degeneration in mice which you can see

in that picture in the lower right hand

corner the left-hand retina is a normal

retina and the one on the right is the

one with the rd8 mutation you can see

that retinal degeneration they're all

c57 black 6 and

substrains are homozygous for the RDA

mutation but the c57 black 6 jameise are

wild type for that mutation so that

mutation happened after the mice had

gone to nih in 1951 and happened in that

colony and now all other derivatives or

other c fat these simplex and sub

strains are then homozygous for that

mutation and the consequences of that

could be in the interpreting of genes or

data as where you're looking at either

phenotypes that involve site or involve

neurobiology phenotypic analysis of

genes that are implicated in cognitive

function like behavioral tests like the

Morris water maze which requires visual

cues if you have c57 black 6 and mice

they're not they're going to have vision

problems which means they're going to

respond differently due to that mutation

not necessarily due to any treatment or

other mutations or transients that you

have and this can impact your areas of

research like Alzheimer's or autism Down

syndrome red sindermann

and a whole host of other

neurodegenerative disorders so it's

really critical that you understand that

if you are using those mice because that

could be an issue again in properly

setting up your experiments in

interpreting your data and this has

really come to be a sort of a major

issue with the retinal degeneration in

the black six and mice as there's this

international knockout mouse consortium

or ikm C which is an international

project to knock out all the protein

coding genes in the mouse but it just so

happens that the ESL lines used to

create all these mice were from a c57

black 6 and substrain meaning that all

of these mice are homozygous for that

rd8 mutation and this again is a

combination of strains from the knockout

mouse project in the US the comp the

European conditional Mouse mutagenesis

project or you come and then also the

the Norcom group in Canada and then

there's also the Texas AM it's a to

traditional medicine ticking which is a

separate thing but where they're making

gene traps but again they're all in c57

black 6 and sub strains which means c57

black 6 and becomes the better control

but we'll have the retinal degenerations

phenotype so if you're looking at

behavior that were quite a vision it's

going to be a an issue in addition to

generating all of these

knockout mice the ikm see also has

designated three different phenotyping

centers to start collecting phenotypic

information for all of these different

mutations and have looked at

reproducibility and they did look at

comparison the black 6j in the black 6n

and looking at a whole series of data as

you can see here this is just a subset

and the way this data is indicated is

any of those rectangles here that are in

red indicates that the black 6n have a

higher response than the black 6j and

the the darkness of the red indicates

then a higher difference the rectangles

in green indicate that the black 6n has

a lower value in the black 6j and again

the the darkness of the of the green

color indicates the the strength of that

decrease and when they and they also

looked at males and females for both in

here you can see all this data that's

comparing it and it's relatively very

well consistent between the three

different phenotypic centers that are

being used in that project and some of

these data that has been collected here

isn't just an example of an

immunological difference where the black

6j females actually show greater

susceptibility to infection with

Listeria then any of the other the black

6j males or the black 6n mice altogether

and then you can also see that the black

6n mice show a significant

pro-inflammatory response three days

after infection where they have

significantly higher expression levels

of IL 6 IP 10 and CCL 2 and another

example out of this data set is the

black 6j mice looking at which have a

higher response with delayed type

hypersensitivity which is a experimental

protocol where you sensitize the mice by

challenging them with DNF B on the

stomach and then you come back and

challenge them with it on the ear and

what will happen is they'll react to

that by ear swelling or so they'll get

inflammation in the ear and then you can

just measure the ear swelling as a

physical measurement and it is a t-cell

mediated response and what you see with

this is that the black 6j males and

females significantly have a

significantly greater inflammatory

response than the black 6 and males or females

females

and then also one can look at there's a

lot of information known about the

genetic background in terms of mutations

and snips of the black sixth chain and

black six and in there multiple of those

mutations have been identified as well

as structural variants that that data is

available as well and this knockout

mouse phenotyping project or the comp 2

is to generate in phenotype 2,500

different knockout mouse strains and

it's a it's a collaborative effort

between the three genotyping centers

that Jackson laboratory here in Bar

Harbor Maine is one of those centers it

along with a Baylor College of Medicine

in Houston which also is working with

the Wellcome Trust Sanger Institute and

the MRC and Harwell and the third place

with the phenotyping Center is the UC

Davis which in addition to their

facility is working also with the

Toronto Centre for Fino genomics in

Canada and the Children's Hospital

Oakland Research Institute and the

Charles River lab in Massachusetts so

when we look at the genetic background

of the mice distributed by the Jackson

Laboratory that are honestly 57 black 6

background whether they're knockouts

transgenics or spontaneous induce

mutations nearly 2,000 of those mice are

on a c57 black 6 j background we have

about 70 that are on the black 6n

however we have over a thousand strains

either coming from the comp 2 project or

through youcome that are going to be on

the c57 black 6 n background and so

understanding the differences between

those backgrounds and understanding the

nomenclature differences it's going to

help ensure that you can pick the right

control strain for the strain that

you're getting and that you understand

that there are differences between them

and so of course when you start looking

then with these different sub strains we

have to talk about the controls and how

you choose the right control so if you

have a congenic mouse strain where you

have a mutation or transgenes that's

been back crossed to c57 black 6 whether

it's black 6j or black 6 and for more

than ten generations you can typically

use littermates

as controls if you're doing het by hat

or hep by wild-type or if it's a

transgene hemi by wild-type if any of

those mating schemes and in those cases

you can always use wild-type or

potentially the heterozygous littermates

as controls for the mutant gene or

allele if you have transgenic mice you

would use the non carrier to the

wild-type mice but you could also

potentially use non littermate controls

from the colony so that would be say

mice from other litters that are all

from the same colony if you're breeding

homozygote by homozygote then you could

use the inbred substring as your control

but again you need to make sure you

understand what substring you're

homozygous mutants are on so you can

match the background as close as

possible to make sure you have the best

possible control if you have a mixed

background say a mix of black six jane

black 6n which is actually more common

than you might think and I'll discuss

that in a little bit coming up but it's

not uncommon for people to not either

know what background they're black six

mice are on or potentially to get mice

from one vendor at one time say black 6j

but then the next time maybe somebody's

got a kind of a little discount going on

so they get black 6n the next time and

now they have a mix of both and if they

don't understand their significant

genetic differences between those they

may not be overly concerned but in those

cases really your only options are

litter mates so that would either be

wild-type or heterozygous for the

mutants more non cures for the

transgenes or again your non-letter mate

controls from the colony I'd like to

give one more example so if you remember

back when I was talking about a high-fat

diet and the daityas versus B City I

gave that hypothetical situation I

actually want to show you actual real

data that has been published that ran

into that same conundrum with choosing

controls so these are researchers they

had generated a JMK 2 also known as map

kinase 9 knockout on a c57 black 6

background and what they did is they

then used acetaminophen induced liver

injury was their phenotype they were

looking at so they compared that

knockout to the c57 black 6 jake

controls and what they found is that the

knockout mice had a higher degree of

liver injury indicating that the map

kinase 9 gene product or the protein was

protective of that phenotype which was

turned out to be the opposite of what

the researchers believed was going to

happen with those

come ice so they then looked at compared

then the knockout to the c57 black 6 and

J and they then found that the black 6nj

had a higher amount of acetaminophen

induced liver injury than their knockout

mice so that left him in that situation

where if you compare the knockout to the

black 6j mice that indicates that genus

paddle protective but if you compare it

to the black 6nj that indicates that the

junk to protein is Patou toxic so that

left them in a position where now what

is the best substring for their

experiments then they actually looked at

the genome and they found that it turns

out that their mouse was actually on a

black 6n genetic background not black 6j

therefore the black 6 NJ is the better

control and it looks like then that

protein is a pedal toxic so it's that

protein itself is involved in that

increased acetaminophen induced injury

but again if they didn't choose the

right control or they didn't have done

both and they had just looked at the

black 6j all of their data would be

reported incorrectly so they would have

interpreted that wrong and then

everybody else who may have followed

that research and built on it would have

been doing things incorrectly

potentially so if you are getting any

new Mouse strain whether it's from a

colleague or a collaborator or you know

somebody published a strain and they're

willing to give you mice there's a lot

of information you really want to ask

them so that you can understand the

genetic background of the mice they're

sending you and so that way you can

choose the right controls so you want to

know what strain was used to develop the

strain was there an O site donor

involved what was that was it yes cell

line what es cell line was used any

other strains that might have been

introduced their breeding you know do

they cross it to a career a flip strain

did they cross just some sort of

reporter like a lag 0 GFP reporter or

other mutations did they remove it or

those mutations still there it's a

current breeding scheme are they

bringing up my hat home by home what are

they doing it's nice to know the current

generation been crop has served in any

point if so what generation has been

back crossed to an inbred strain if so

what inbred strain have they confirmed

the genetic background so if they say

the mysimon back

ten times the c57 black 6 J have they

actually confirmed that it really is

well back crossed and so at the Jackson

Laboratory we do have stringent QC

quality control that we then look at the

genetic background of all the strains

that we import and when we do that it's

not uncommon that we find that the the

genetic background of the mouse that we

import is actually not what we are told

in fact almost a 30 I think it's 31

percent of the strains that we look at

are not on the background that we're

told so that's about one out of three I

think something like 16% end up being on

a mix of black 6j and black 6n and so if

that's happening with the strains that

we import just think about what's going

on with the mice that you might get from

a colleague or a collaborator or some

other researcher so it's going to be the

same so if you are getting a new strain

highly recommend that you look at

monitoring the genetic background and at

the Jackson laboratory we do have

services that can help you with that we

do have a genome scanning service that

can distinguish between the c57 black 6

j and the C 50 some black 6 and sub

strains it's 150 markers with markers

distributed evenly throughout the genome

so we can distinguish those two we can

also distinguish whether the mice have

been back Ross safe to c57 black 6 you

know whether it was on a 129 or went

from bob see we have panels that we can

use to distinguish between any two

commonly used inbred mouse strains and

confirm the background and I would

highly recommend if you are getting a

new strain from other people I would

highly recommend you take that time and

look at confirming the genetic

background because it's better to know

upfront that the mice are not the

background you expect them to be so that

you can then plan accordingly whether

alter your controls or potentially

backcross but it's better to know before

and after you get a lot of data

potentially find the problem or you may

never know there's a problem and there

you end up with the wrong control and

again you wouldn't be in a position

where you would interpret your data

incorrectly I mentioned earlier that the

Jackson laboratory we do have two

substrates of c57 black 6 both the black

6j and the black 6 NJ again these are

very well characterized we have them

both at a high health status and

I had talked about data reproducibility

so you know what are we doing here at

the Jackson Laboratory to ensure the

genetic integrity of these two sub

strains and so what we have developed is

a genetic stability program there's a

patented program that we developed here

at the Jackson Laboratory that we use to

diminish cumulative genetic drift and

thereby stabilize the phenotype of those

mice and so what we have for these

strains is a 25-year supply of frozen

embryos from the same generation and

what we do is we thought some of those

embryos to generate our foundation

colony and the foundation mice then

breed for at the most up to five

generations after that point we would

remove them from the shelves again

generate new mice from the what we have

crab preserved during the five

generations of breeding the foundation

we then pull off mice for the expansion

of distribution where those mice breed

it for most ten generations which would

then of course be the mice that you

would be purchasing directly from us so

therefore the mice that you get from us

for either the black 6j or the black 6nj

are at most 15 generations removed from

what we have cryopreserved

now during those generations mutations

will happen but because we keep going

back to the beginning those mutations

are not able to fix or become homozygous

in the colony

therefore the mice that you get from us

today are going to be as genetically

identical to mice that you get five 10

20 years from now thereby were then

maintaining that genetic stability and

then retaining that same phenotype so

again that the phenotype and the

genotype should say stable every year

and we have a lot of resources that we

can't use to support using black six

mice we have the mouse genome database

where we collecting baseline phenotypic

information for a host of commonly used

inbred mouse strains including the c57

black 6 J where we have close to 3,000

different measurements so you can use

that to either get baseline information

for the phenotypes you're interested in

that way you don't have to establish

them yourself in your lab but you just

can validate them there is a whole

genome sequence data available from the

Sanger Institute and the mouse genomes

project the c57 black 6 J was the

original Mouse genome that

originally sequins but the black 6nj has

been sequenced with this mouse genome

project therefore you can search for

mutations between those we also have a

lot of precondition mice for the strains

where we including a strep disease and

induced diabetes or diet induced obese

where you can get mice black 6j mice

that are already fed a high-fat diet as

well as aged mice as well and then both

of these strains are available at a very

high health status with no additional

charges for that it's all all the same

you know I just want to make sure you

guys are aware again that's really

critical to understand the phenotypes of

the stranger using and you can use the

mouse genome database or MPD to get some

of that baseline phenotype here's just

an example of plasma glucose levels

after a four hour fast and looking at 43

different Mouse trains the arrow is

pointing to the black 6j so you can use

of course the MPD database of using any

commonly used in bran strain not just

c57 black 6 but if you are using black 6

which most of you certainly are you can

find baseline data in that database

which can then have an idea what to expect

expect

prior to any treatment or mutation and

so really you need to really consider

that your experimental model is one of

your most important reagents you know

really choose wisely and really

understand the inbred strain you're

using as well as the sub strain the more

you know the less problems you'll run

into because you you really understand

that always really critical to use

proper nomenclature so if you're using

black 6 mice indicate you are using

black 6j are using black 6 NJ black 6 n

c RL it's really critical to really use

that that way anybody else who reads

that publication will know what strain

you used and then again you know

minimize genetic drift either get your

mice from a respected vendor that is

working to minimize genetic drift or if

you've got a mutation or transient on a

black 6 8j inbred background you want a

back cross to the black 6j every five to

ten generations to keep your mice as

similar as to your controls as possible

make sure you're well educated and

really establishing a good quality

control culture because really the

better you understand and the better

quality control you take then the less

mice you'll have to use and really it's

really good

does result in using reduced animals so

the less mice you have to use the less

resources and timer involved in that

will improve your research program so

again just to summarize there are many

different genetically and phenotypically

unique sub strains of c57 black 6 that

have developed over time and they're

going to continue to develop as time

goes on and really knowing and

understanding the black 6 sub thing that

you're working with is really critical

to choosing the proper controls and

proper data interpretation looking at

the phenotypes between black 6 sub

drains can also allow for identification

of unique modifier alleles so while you

may think that the different sub strains

might be a headache to have to pay

attention to they may offer really

really good experimental models to look

at those phenotypes that are different

that the Jackson Laboratory we're using

our genetic stability program to limit

genetic drift to stabilize those

phenotypes over time this presentation

was made possible with your support

Jack's Meissen Services is dedicated to

providing you with quality resources and

flexible solutions and innovative

technologies and unsurpassed expertise

to support all aspects of your research

and realize that every dollar that you

do spend at Jack's mics and services

does support our shared research goals

and sustains all these valuable mouse

models and resources that are only

available from the Jackson Laboratory we

do offer a host of services that might

be helpful to you asides just having the

specialty strains we also do on basic

custom and complicated complex breeding

capabilities so if you're having trouble

breeding or you wish we can breed creole

ox mice together for you and either send

you the breeding pairs or maintain

cohorts for you here as well as doing

compound efficacy testing and genome

scanning as well and with that again

thank you for attending this

presentation and if you do have any

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