Question: (Closed) Poor PCR product
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gravatar for loly.pearl86
2.7 years ago by
loly.pearl8630
Australia
loly.pearl8630 wrote:

Hi everyone,

Actually, i am working on gene expression project and aimed to use RT-PCR and NGS methods. The NGS method work well BUT the PCR product for RT-PCR method was poor.

I designed the primers using primer-BLAST and then run PCR to check the presence and absence of the product. I run the gel but there was no products/no amplifications. I tried other primers but still no products. My target sequences have a lots of low complexity regions, and based on what i read the low complexity regions consume the PCR product and caused the failure PCR (no products). So I concentrated on the NGS method only instead of using RT-PCR. My question is ..is this enough to justify the reason of excluded the RT-PCR method cause of the low complexity regions which is led to the failure PCR or i should try other primers until its work ?? Or there is another possible reasons to get this failure PCR?

Thanks

gene • 527 views
ADD COMMENTlink written 2.7 years ago by loly.pearl8630

Hello loly.pearl86!

We believe that this post does not fit the main topic of this site.

Not related to bioinformatics

For this reason we have closed your question. This allows us to keep the site focused on the topics that the community can help with.

If you disagree please tell us why in a reply below, we'll be happy to talk about it.

Cheers!

ADD REPLYlink written 2.7 years ago by WouterDeCoster44k

Please post this over at SeqAnswers.com.

ADD REPLYlink written 2.7 years ago by genomax91k
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