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

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==Size exclusion filter purification==
==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===
==Folded samples BM_2_1565 (23.8.2011) and BM_2_5Dh3ml (23.8.2011) were filtered in the following manner==




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   - 3 x timed filtered buffer discarded
   - 3 x timed filtered buffer discarded


===Ran a 50 ml 2% Agarose-Gel with EtBr===
==Ran a 50 ml 2% Agarose-Gel with EtBr==


   - 1kb ladder
   - 1kb ladder

Revision as of 04:06, 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!!!