Internal Control RNA & RT-PCR - (Jul/06/2005 )
I'm doing Real Time PCR for viral RNA detection.
Do you have any idea on how making an Internal Control (heterologue RNA sequence?), that can be added indifferently to extraction or to PCR and that is amplified togheter with my viral target in a multiplex?
I am working with multiplex Real-time PCR but for DNA. For Internal Control I have used a conserved gene like 16srRNA which is present in multiple copies in the organism. But this can sometimes suppress the amplification of the target gene which is present in less number of copies.
I am not familiar with the viral genome. But you can use any gene that is present in the virus and is conserved as the internal control.