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generating cDNA from human brain RNA


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#1 Fluries

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Posted 05 January 2011 - 05:59 PM

Hi all,

I have extracted some RNA from human brain samples and want to generate cDNA from this. I prepared cDNA using random hexamers, oligo(dT) and gene specific primers. Next I try to amplify the genes of my interest, but do not get any product after PCR. All i see is the low molecualr weight primers or primer dimers.

For gene specific primers I used 2 ul of 1uM antisence primer in a 20 ul reaction volume and i do RT for 50 min at 55 C.

The genes of my interest range in size from about 1Kb to about 6 Kb.

Any suggestions as to what I am doing wrong or what else I can try.

Regards

#2 ElHo

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Posted 06 January 2011 - 01:41 AM

Did you check the quality of your cDNA by amplifying a housekeeping gene or any other gene already established in your lab?




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