Biomod/2011/TUM/TNT/LabbookA/2011/09/06: Difference between revisions

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Ask Marty about buffer and exact procedure!!!
Ask Marty about buffer and exact procedure!!!


==Gel picture==
insert jpeg!!!
looks like there are residual staples in solution and the leading band is slower as the scaffold. Marty said this is normal, when purified samples are put on a gel.




Revision as of 04:14, 6 September 2011

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Size exclusion filter purification

Folded samples BM_2_1565 (23.8.2011) and BM_2_5Dh3ml (23.8.2011) were filtered in the following manner

  • 50 µl from both samples were mixed and put into a Amicon filter
  • 400 µl buffer added
  • spinned for 5 minutes at 14000 rcf
  • 400 µl buffer added
  • spinned for 5 minutes at 14000 rcf
  • 400 µl buffer added
  • spinned for 5 minutes at 14000 rcf
  • turned the filter and sinned the purified sample into a new Epi for 2 minutes at 1000 rcf
  • 1 x and 2 x filtered buffer filled into separate Epi (the buffer, which came through the filter; if purification with spin columns works, this should only contain staples)
  • 3 x timed filtered buffer discarded

Ran a 50 ml 2% Agarose-Gel with EtBr

  • 1kb ladder
  • p7560
  • filtered BM_2
  • BM_2_1565
  • BM_2_5Dh3ml
  • 1x filtered buffer
  • 2x filtered buffer

Ask Marty about buffer and exact procedure!!!

Gel picture

insert jpeg!!!

looks like there are residual staples in solution and the leading band is slower as the scaffold. Marty said this is normal, when purified samples are put on a gel.


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