Long RT-PCR

Jim Owens jow at helix.nih.gov
Wed Jun 1 07:41:21 EST 1994


In article <1994May31.155954.1149 at cathy.ijs.si> Kristina Gruden,
Kristina.Gruden at ijs.si writes:
>I would like to get a 2,6kb long RT-PCR product. I've succesfully
produced
>a 500bp product but have no expirience with products that long. I know
that
>this lenght shouldn't be a problem if performing only PCR. But I read
somewhere
>that longer RT-PCR amplification can make some troubles. If you have any 
>expirience with this problem I will be glad to hear from you whether you 
>did it by standard procedure or you used some special modifications.

I'm interested, too.  I have been performing RT-PCR on translocated c-myc
loci in mice and have found repeatedly evidence that the cDNA synthesis
has made longer products than I can PCR.  That is, I can amplify several
segments less than a kilobase but cannot amplify the full 3-4 kilobase
cDNA.

Good luck,

Jim Owens



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