由买买提看人间百态

boards

本页内容为未名空间相应帖子的节选和存档,一周内的贴子最多显示50字,超过一周显示500字 访问原贴
Biology版 - 基于Li-COR的southern blot,求帮助
相关主题
问个单链DNA连接的问题Trouble-shooting of Southern Blot
Re: [Question 1] About microarrayRe: 求助: Southern Hybridization with olig
请推荐做In situ hybridization的公司各位觉得taqman probe的specificity如何
Re: help on ligation problem!有啥公司能给做in situ hybridization 用的plasmid
菜鸟定向克隆求助,谢谢~RNA-seq测的也不是绝对表达值吧
digest 100ug plasmid DNA 最小体积多少啊?RNA in situ hybridization请教
Histone DNA bindingThe best way to keep healthy in winter is to turn heater down
有做antibody engineering的筒子吗?有一个digestion的问题
相关话题的讨论汇总
话题: probe话题: dna话题: li话题: cor
进入Biology版参与讨论
1 (共1页)
R****n
发帖数: 708
1
最近做一个genomic feature需要用20bp左右的probe hybridize digested genomic
DNA.有几个问题
1)overnight transfer DNA 到 nitro membrane之后,压扁的胶里还是有 DNA smear,
是不是没有transfer成功,还是有残留?
2)LI-COR的protocol是对PCR probe优化的 (65C hydridization, 50C wash),
https://app.boxenterprise.net/s/swc2yyeseopg608jlyiuqe8qs9t33rb9
怎么优化hybridization温度?这篇文章提了一下.关键这个hybrization溶液里有
Dextron Sulfate 和 近 700mM的Na+.算算有68C了。。。
http://www.alliot.fr/BIO/PDF/SouthernBlot-.pdf
short oligonucleotide probe is used (15–25 nucleotides in
length),then the hybridization step is usually carried out
under conditions of low stringency (typically at a
temperature several degrees below the calculated Tm for
the desired hybrid),so that all potential hybrids,including
nonspecific ones,are able to form。 Specificity is then
achieved by a series of washes at increasing temperatures so
that,hopefully,only the desired hybrid remains at the end
of the procedure.
3)probe是biotin labelled.我看了一下以前的讨论,牛奶肯定不能用来bloxk。 1%
BSA 应该没问题吧?
1 (共1页)
进入Biology版参与讨论
相关主题
有一个digestion的问题菜鸟定向克隆求助,谢谢~
How can I activate the old PVDF WB membranedigest 100ug plasmid DNA 最小体积多少啊?
求分离x chromosome的ideaHistone DNA binding
双酶切奇怪条带有做antibody engineering的筒子吗?
问个单链DNA连接的问题Trouble-shooting of Southern Blot
Re: [Question 1] About microarrayRe: 求助: Southern Hybridization with olig
请推荐做In situ hybridization的公司各位觉得taqman probe的specificity如何
Re: help on ligation problem!有啥公司能给做in situ hybridization 用的plasmid
相关话题的讨论汇总
话题: probe话题: dna话题: li话题: cor