I would like to know more about presence of sugars, or in fact all carbohydrates in PCR.
What are their effects? What should we do to remove them from the PCR reaction?
Heparin (a mucopolysaccharide) severely inhibits PCR. It is sometimes difficult to do PCR on plant material on plants unless you add CTAB (cetyltrimethyl ammonium bromide) to the buffer to remove polysaccharides and other interfering substances (you can look up a protocol on the internet). I once know a person who was trying to do PCR from snails (which produce lots of mucous, and hence, mucopolysaccharides), and had trouble until he added CTAB to the extraction buffer. If you are having trouble with PCR using DNA from plants or organisms with lots of mucous, you could try adding CTAB to your extraction buffer. Otherwise, I do not think most carbohydrates have much effect on PCR.
Heparin (a mucopolysaccharide) severely inhibits PCR. It is sometimes difficult to do PCR on plant material on plants unless you add CTAB (cetyltrimethyl ammonium bromide) to the buffer to remove polysaccharides and other interfering substances (you can look up a protocol on the internet). I once know a person who was trying to do PCR from snails (which produce lots of mucous, and hence, mucopolysaccharides), and had trouble until he added CTAB to the extraction buffer. If you are having trouble with PCR using DNA from plants or organisms with lots of mucous, you could try adding CTAB to your extraction buffer. Otherwise, I do not think most carbohydrates have much effect on PCR.
Glucose-6-phosphatase (G6Pase, Heparin and undigested proetoglycans hinder qunatification using real-time PCR (targeting 16S rDNA) .
Christopher is right, you need to purify your template, and CTAB is a good way to eliminate polysaccaride. I work with gelatinous animals (a lot of mucus), and I use CTAB phenol/chloroform extraction. The results are perfect!
Thanks Dr Theodorakis.thanks for your full answer.
thanks dear Dr Ravi Kant Upadhyay and Dr Aglieri.
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