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cDNA Forum cDNA Forum. Discuss complementary DNA synthesis, cDNA libraries, and cDNA cloning in this forum.


cDNA Vs DNA

cDNA Vs DNA - cDNA Forum

cDNA Vs DNA - cDNA Forum. Discuss complementary DNA synthesis, cDNA libraries, and cDNA cloning in this forum.


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Old 12-13-2008, 02:45 PM
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Default cDNA Vs DNA



I have been trying to understand some cDNA extraction techniques, but could someone explain me what makes a particular set of primers (forward and reverse) recognize and amplify cDNA, and not DNA, in the same conditions?
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Old 12-15-2008, 03:34 PM
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Default Re: cDNA Vs DNA

If a primer recognizes cDNA, it will generally recognize genomic DNA as the chemistry of the two is identical. You get specific cDNA amplification if:
1) you get rid of the genomic DNA, for example using DNase at the RNA isolation step or selectively precipitating RNA with LiCl
2) The primer sits directly over a splice site, so that not enough of the primer recognizes either genomic DNA sequence to bind well
3) The primer pair spans a splice site, so that the product in the genomic DNA is much longer than in the cDNA, and the PCR conditions are such that the longer product doesn't form
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