In my experiment after ligation process It is required to block the 3' end of oligonucleotide from polymerase extension.
Normally after the ligation heating will deactivate the ligation process while there would be some possibility of extension during PCR, in case is there any enzyme that can block the 3' end?
dear sir, what would be the application of your pcr product. if you add the enzyme alkaline phoshphatase it will remove the phoshphate group from 5' end thereby subsequent ligation can be prevented. Are you trying to prevent the ligation of your pcr product?
Dear Hariharan,
Thank you for your attention. In fact we are working some new method in ligation process and in-order to prove the efficiency compere to conventional methods inhibiting any ligation during PCR is required, so I wonder if there would be any possibility of blocking only 3' end terminal not 5' terminal
Here is the steps
1) Phosphorylation
2) Ligation
3) deactivating of ligation by heating
4) in this step I want to block the 3' end terminal to prevent any ligation in PCR
5) PCR
Dear Andrei,
Thank you for your attention, Does it effect just only on 3' terminal?
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