K**R
发帖数: 193 | 1 想请教，northernblotting 探针一般设计多长呢？ 有地说50bp 有的说1KB。。。 | B******o
发帖数: 496 | 2 Your information is too limited. What is your target? mRNA or small RNAs?
How about your probe, DNA or RNA?
For small RNAs, complementary DNA probe is more than enough.
For mRNAs, people used in vitro transcription to generate ~500nt DNA probe.
【在 K**R 的大作中提到】
: 想请教，northernblotting 探针一般设计多长呢？ 有地说50bp 有的说1KB。。。
| K**R
发帖数: 193 | 3 谢谢您的回复，
我是为了测试alternative splicing。 我们最近发现一个基因alternative splicing
在发育和成熟组织中似乎存在，想证实是否是western 非特异还是确实是alternative
splicing。
想设计一段包含所有潜在的splicing，就是shared的区域。然后northern 测试是不是
确实有不同大小拼接。
probe应该是cdna 为模板和mRNA杂交
谢谢您
.
【在 B******o 的大作中提到】
: Your information is too limited. What is your target? mRNA or small RNAs?
: How about your probe, DNA or RNA?
: For small RNAs, complementary DNA probe is more than enough.
: For mRNAs, people used in vitro transcription to generate ~500nt DNA probe.
| g*********5
发帖数: 2533 | | B******o
发帖数: 496 | 5 then in vitro transcription will be your choice to generate northern probes.
However, I agree with the other suggestion that you should do RT-PCR first.
Remember to include DNase treatment.
在 KSOR (KK) 的大作中提到: 】
splicing
alternative |
|
