What is the best method for cDNA synthesis from miRNA?

Hello,

I have a linearized plasmid (digested and cleaned by phenol/chloroform precipitation) that has been stored in water at minus 20 deg for two weeks. The plasmid is intended to be used as template for mRNA synthesis and I am wondering if I should worry about degradation/hydrolysis by this time. And generally, what would be the maximal storage time for linearized plasmids in water at minus 20?

Thank you for your help!

Zohar

I am working on RNA seq data (For banana plant) and we predicted that Intron-retention ( Alternative splicing) is predominately occurred. However, i need to extract intronic sequences from these large RNA seq data.Actually i tried with a alternative variants of a gene from RNA seq to blast with banana CDS of that gene. But not able to find much differences. Moreover i do not aware of the sequence characteristics of introns. Any possibe help in this regards? Thanks in advance

I'm searching for a tool that helps me to find promoter of miR-455-5p. Is there any suggestion?

Hi,

I'm going to do DNA sequencing from mRNA directly extracted from blood's patient.

Is there anyone who has used from QIAGEN Kit ?

I am looking to run some qRTs on some isolated miRNA. I've used a kit to isolate the small RNAs from the total RNA. Is there a need for me to get a cDNA synthesis specific for miRNAs? If so, why? How are the small RNAs different from the larger RNAs for RT? 

Additionally, will I need sybergreen/taqman kits designed for miRNA detection as well?

Thanks!

- the IRCs rise one to three cycles earlier then all the other samples with every primer