Formal Lab Session - 6th October 2009
Today we did B. subtilis transformations using our ptotocol. We prepared the LB+ CHL(Chloramphenicol) plates since we will transform the cells with pgfp-rrnb. We used 5ml 2.5 mg/ml stock of CHL, however it should have been 2 ml since the final concentration should have been 5ug/ml CHL.
We did miniprep to look for CotC+ pmutin4 colonies. We used 12 ON cultures. Finally we did a restriction digest with HindIII and BamHI. But we could not find a right colony again. Although the transformations seemed worked we had colonies on both "backbnone+ no ligase + no insert" and "backbnone+ ligase + no insert". Our transformations dd not have the ligated product!
Digest:
H20 11ul
Buffer 2ul
DNa 5ul
HindIII 1ul
BamHI 1ul
Control Digest:
H20 13ul
Buffer 2ul
uncut pmutin4 2ul
HindIII 1ul
BamHI 1ul
The bands were pmutin4 were not very clear and we decided to do a midiprep for pmutin4 again.
July
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August
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