Designing primers with ESTs
Posted 27 February 2010 - 04:52 PM
I have decided to design primers using EST sequences since the species (sus scrofa) I am involved with have less mRNA data in Genebank. I am doing RT-PCR for my microarray validation. What are the pit falls and risks of designing primers using ESTs. Can I cite them like mRNA sequences? Please let me know what you all think. thanks in advance.
Posted 27 February 2010 - 06:37 PM
Posted 28 February 2010 - 12:41 AM
it is totally feaisble to design primers based on EST sequences if there is no known mRNA sequence and to cite the sequence in your papers. The more EST sequence alignment for the region of your interest, the safer that the region is a true mRNA sequence.
Thanks for the reply. appreciated.