when I constructed library for Next-generation sequencing, the efficience of adapter ligation was low, How can I improve it? chage the enzyme or the condition of reaction.
Hi Fei,
What type of ligation do you do? Are they blunt or cohesive? in the first case they are less effective and you can therefore consider ligation after a step "+ A". Make sure that the ratio of adapter / fragment is approximately 20 (in molarity). You can also increase the ligation time.
Best regards
Cédric
Mariac Cedric Thanks for your advice. in the other wise, there is a limitation of end-jioing efficience for T4 DNA ligase using in NGS library constructed. Some one use a engineered DNA ligases that fused T4 DNA ligase to different DNA-binding proteins, and it improved the efficience of adapter ligation significantly . Is there any commercial kit using the fusion T4 DNA ligase？
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