RT-Pcr without RNA isolation? - (Oct/17/2007 )

Hi there!

Does anyone have a protocol for RT-PCR from cell culture cells without having to isolate the RNA first and degrading the DNA?

Thanks in advance

Stardust

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There are diverse products from different manufacturers that offer "one tube" RT-PCR reactions, even starting from whole cells...

E.g. :

http://www1.qiagen.com/Products/Pcr/Quanti...ellcDNAKit.aspx
"For high-speed preparation of cDNA without RNA purification for use in real-time RT-PCR"

or

http://www1.qiagen.com/products/pcr/FastLa...BRGreenKit.aspx
"For SYBR Green based, real-time, one-step RT-PCR direct from cells without RNA purification"


Mike

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The problem is, we don't have enough money to buy the kits...that's why i'm looking for a "homemade" option...

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I understand your condition as my group is also not a rich group. Then, forget about kits, back to the conventional method. Or you may want to study those company product in detail and see if you can make them up by yourself.

Good luck.

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