RT-PCR primer design
Posted 15 October 2004 - 05:43 AM
Can you help me?
I need of a program to detect in mRNA sequence the exon junctions. Becouse I need to design primers that donīt regonize genomic DNA.
Posted 15 October 2004 - 04:13 PM
To answer your question about RT-PCR primer design, you can read this post:
Edited by sage, 15 October 2004 - 04:14 PM.
Posted 27 October 2004 - 06:13 PM
To design your primers for RT-PCR you can do a search of the ORF of your interest gene. You can select the codifing region and design your primers uppon that sequence. I have had very good results doing this.
Posted 27 October 2004 - 06:47 PM
I could not agree with you. You have to check the genomic structure of your gene to make sure the RT-PCR primers either span at least one intron or bridge exon-exon junction. You can first design primers using the coding sequence, then check if the amplified region contains any introns.
Posted 28 October 2004 - 10:13 AM
I think you're right. I didnīt considered the exon-intron junction. Thanks for the corrigendum!