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troubles to make a 3 ways DNA ligation

troubles to make a 3 ways DNA ligation - DNA Techniques

troubles to make a 3 ways DNA ligation - Post questions and discuss DNA techniques and protocols such as DNA extraction, PCR, and the study of DNA-binding proteins.


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Old 09-15-2010, 12:00 AM
Pipette Filler
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Default troubles to make a 3 ways DNA ligation



I have troubles to make a 3 ways DNA ligation

My setup is

1] Vector pEt 15b digested with Nde1/Xho1 and dephosphorylated and cleaned up with qiagen kit
2] Insert 1, 210 bp from pcr product digested with Nde1/Bamh1 and cleaned up with qiagen kit
3] Insert 2, 210 bp from pcr product digested with BglII/Xho1 and cleaned up with qiagen kit too
I have tried a molar ratio vector/insert 1/3 without success
Have you some advices for me
Thanks
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