h***9
发帖数: 662 | |
D***s
发帖数: 5613 | 2 用过invitrogen(Life technology)的5' RACE System for Rapid Amplification of
cDNA Ends, Version 2.0，感觉还可以，GSP引物设计比较重要，最好多设计几条
【在 h***9 的大作中提到】
: clontech的怎样？
: 谢谢
|
p******b
发帖数: 379 | |
h***9
发帖数: 662 | 4 谢谢Diors 和polycomb。
这个kit,做3'race好使不？
【在 p******b 的大作中提到】
: http://www.lifetechnologies.com/order/catalog/product/AM1700M
: 我用的是这个， 很好用
: FirstChoice® RLM-RACE Kit
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p******b
发帖数: 379 | 5 FirstChoice® RLM-RACE Kit 我3'和 5'都试过， 都挺好用的，goodluck |
h***9
发帖数: 662 | 6 Thank you!
【在 p******b 的大作中提到】
: FirstChoice® RLM-RACE Kit 我3'和 5'都试过， 都挺好用的，goodluck
|
l**********1
发帖数: 5204 | 7 RE LZ
only your 3’RACE does not work with that kit.
i mena
if your target mRNA its 3' UTR that is super long (>4kp) or before its ROF
stop codon there is/are high GC rich region/s, you can try another method:
一句话就是用内切酶对应的人工引物锚定: 3'端 做人工锚定引物PCR
mRNA with a primer (3'-poly
(dT)17 with an anchor sequence at its 5'end, then qRTPCR then PCR sub
cloning... sequencing.
References:
HTTP double dot //genome.cshlp.org/content/4/1/19.full.pdf
http://www.ncbi.nlm.nih.gov/pmc/articles
Or even up/down stream of circled mtDNA can be PCRed with above method:
http://www.ncbi.nlm.nih.gov/pubmed/19605532
more details pls refer attached figure.
or pls go to
http://www.ipv6.weiming.info/zhuti/Biology/31635707/ |
