at Qiagen therascreen pyro kit, there a dNTPs. but in there, dATP is modified, dATP alpha S. So I found why uses dATP alpha S, but there's no answer...
why uses dATP alpha S in pyrosequensing?
Hi Kyungson
The dATPαS is used as a substitute for the dATP since it is efficiently used by the DNA polymerase, but not recognized by the luciferase.
Good luck,
Silvia
Hi there!
You already have got your answer, but I'll just like to elaborate for easy understanding. Pyrosequencing counts on the number of release of pyrophosphate after addition of dNMP to the new strand by DNAP. But the read out is through the activity of Luciferase, which needs ATP and there is also the release of pyrophosphate. So, ATP utilised by luciferase will also relase the pyrophoshate and give the read out of ATP incorporation in the DNA sequence, which will be misleading. To differentiate the dATP incorporation against a T (Thymidine) in the sequence and ATP required by Luciferase for its activity, ATPαS is used.
Hope it helps!
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