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Biology版 - Re: 关于genotyping 的几个问题.
相关主题
genotyping 问题求助有没有什么基因和ES cell的maintenance无关,而在器官(胚层)发生中有作用
请教如何保持transgene mice。genotyping.
endogenous protein ip 时,用antibody crosslinked beads incubate cell lysis 多做转基因老鼠的问题
siRNA初级问题请教不同背景小鼠杂交的纯化问题
总是在trouble shooting两个cre的transgenic mice cross有问题
how to over-express a gene?做了cell fractionation以后, nucleus里面啥都没了。。。
一个蛋白,染色可以,western检测不出来是怎么回事生物技术类杂志
新手请教 'Genotype-phenotype correlation’paper needed
相关话题的讨论汇总
话题: transgenic话题: run话题: bands话题: sizes话题: depending
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1 (共1页)
b*********t
发帖数: 877
1

depending on the sizes of the bands (usually two). if they are both big and
close in size, then you will have to run it really slow for a long time (i
have one 11kb/9kb southern that i run at 40V for 10 hrs!); otherwise, you
can run it much faster (70V, 3 hrs.)
this should have been determined BEFORE you make your transgenic mice.
you should be able to predict the sizes/conditions based on your original
constructs.
depending on your probe. usually two bands: one endogenous, one transgenic.
good
M****e
发帖数: 70
2
refer to NAR 1991 19:42 P.W.Laird et al
i have tried to optimize the condition and it works pretty
well for me. basically, cut ~5mm tail when the mickeys are
about 14-d-old (do not forget ear tag), if you worry about
cross-contamination, clean scissors with 70% ethanol and
flame, otherwise, i typically clean it after collecting 5
samples. digest the tail in a tightly sealed epp. with 500ul
lysis buffer, 5ul proteinase K (10mg/ml). rotate O/N at 55C,
though 6hrs is pretty much sufficient. spin 10
1 (共1页)
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相关主题
paper needed总是在trouble shooting
求推荐:Mouse DNA microinjection service/转基因老鼠注射how to over-express a gene?
求做bac transgenic contruct的公司一个蛋白,染色可以,western检测不出来是怎么回事
大片段DNA克隆新手请教 'Genotype-phenotype correlation’
genotyping 问题求助有没有什么基因和ES cell的maintenance无关,而在器官(胚层)发生中有作用
请教如何保持transgene mice。genotyping.
endogenous protein ip 时,用antibody crosslinked beads incubate cell lysis 多做转基因老鼠的问题
siRNA初级问题请教不同背景小鼠杂交的纯化问题
相关话题的讨论汇总
话题: transgenic话题: run话题: bands话题: sizes话题: depending