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Team:Tsinghua/Notebook/10 July 2010
From 2010.igem.org
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Digest the four fragments with restriction enzyme separately except the SalI and EcoRI at 37℃ for more than 3h. Ligate the four fragments at 25℃ for an hour. | Digest the four fragments with restriction enzyme separately except the SalI and EcoRI at 37℃ for more than 3h. Ligate the four fragments at 25℃ for an hour. | ||
Latest revision as of 06:49, 25 October 2010
Module I, group 2(b)
Digest the four fragments with restriction enzyme separately except the SalI and EcoRI at 37℃ for more than 3h. Ligate the four fragments at 25℃ for an hour.
Ligation system:
H2O 22.5μl 10×buffer 5μl fragments 5μl each T4 ligase 2.5μl Total 50μl
Run a gel.
Result
There isn’t any ligation product exists… Even though it exists, the concentration is not high enough to be detected. We’ll improve the ligation protocol!
