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Old 06-16-2009, 11:35 AM
Abo-Ogiala, Atef
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I did the PCR reaction for one of Arabidopsis mutants to verify the T-DNA
insertion and the results are a bit strange. I mean I should get from the
first generation of the mutant at least one hetero plant but that is not
happened. The whole plants were wild type. So it could be there is no T-DNA
insertion or what? Another question concerning cDNA, I tested the cDNA with
the specific primers of my gene (Lp+Rp) and also with (Rp+LB) and I got
results in both reactions, however I got the RNA from wildtype plants. For
that I have a doubt that cDNA could be contaminated or I have no idea?

Best regards

Atef Abo-Ogiala

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Old 06-17-2009, 08:22 PM
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On Jun 16, 7:35*am, "Abo-Ogiala, Atef" <[Only registered users see links. ]-
goettingen.de> wrote:

Use the SIGnAL T-DNA Express tool to find the T-DNA orientation for
your mutants. ([Only registered users see links. ]). As
recommendation avoid the use of LBb1 primer for Salk mutants, instead
use the LBb1.3 or LBa1 primer. Regarding to your PCR on cDNA just
repeat it but extract RNA from Arabidopsis WT plants grow up by
another person, if no possible grow a new batch of WT plants.

Hope this help!

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