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Team:Tsinghua/Notebook/9 August 2010
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| - | == Module I, | + | == Module I, group 2(b) == |
Measure the concentration of the plasmids purified last night. Double digest to ensure the positive clone. | Measure the concentration of the plasmids purified last night. Double digest to ensure the positive clone. | ||
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37℃ 1h, then | 37℃ 1h, then | ||
+ buffer Tango 2.5μl | + buffer Tango 2.5μl | ||
| - | + EcoRI | + | + EcoRI 1μl |
| - | Total | + | Total 23.5μl |
Run a gel after the digestion is finished. | Run a gel after the digestion is finished. | ||
Latest revision as of 06:54, 25 October 2010
Module I, group 2(b)
Measure the concentration of the plasmids purified last night. Double digest to ensure the positive clone.
Double digestion system:
P+E:
H2O 6-8μl buffer Tango 4μl P+E 4-6μl SalI 2μl HindIII 2μl Total 20μl
P+C:
H2O 6-11μl buffer Tango 2μl P+C 4-9μl KpnI 3μl Total 20μl 37℃ 1h, then + buffer Tango 2.5μl + EcoRI 1μl Total 23.5μl
Run a gel after the digestion is finished.
result
We use the 0.8% agarose gel running first and it is quite un-clear around where 700bp band should be. So we run a 1.2% gel but it seemed that there is no positive result among the 12 clone.
