2 replies to this topic

[gopstoro2000]

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Dear all,
Does the commercial PCR products such as the ones from Sigma aldrich , not have the 5' phosphate ends? Its a major problem since i am going for blunt end ligation of PCR product directly and I will ve to know if i ve to do CIPPing of the vector.
With regards
Gopinath.M

[gopstoro2000]

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Dear all
I have myself found the answer for this
The commercially synthesized primers come without a 5'phosphate ends(primers from organisations such as Sigma aldrich) and under special circumstances they also provide primers with 5' phosphate group at an extra cost.
Hope this will be useful for others
with regards
gopinath.M

[vgupta]

You don't need to buy phosphorylated primers. By end filling you automatically generate 5' P. You can not clone PCR product directly as it has poly A overhangs. You need to do end filling