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My question/comment concerns the use of Heparin in the Polysome...
AG
Agata Giallongo Sep 5, 2013
My question/comment concerns the use of Heparin in the Polysome Extraction Buffer (PEB):
heparin is not mentioned in the material and methods section of the full-length paper (see reference)where Rnasin 500U/ml) seems to have been routinely used.
It is known that heparin m Read more
heparin is not mentioned in the material and methods section of the full-length paper (see reference)where Rnasin 500U/ml) seems to have been routinely used.
It is known that heparin m Read more
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HAILONG ZHANG Author Answered Sep 10, 2013
Neoletix Lab, Inc
Hi,Agata, it's a good question. but you don't worry about the downstream applications of RT,RT-PCR,qPCR,because RNA will be precipitated during RNA extraction(step:"21"),and all solutions will be washed away totally(step 24-27). RNA will be very pure and without any heparin any longer.
We need heparin during cell lysis, because the DNA binding sites on RNA polymerase can be occupied by heparin, preventing the polymerase from binding to promoter DNA.
We need heparin during cell lysis, because the DNA binding sites on RNA polymerase can be occupied by heparin, preventing the polymerase from binding to promoter DNA.
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