Team:HokkaidoU Japan/Notebook/September16

(Difference between revisions)

Revision as of 07:42, 19 September 2010

1-14K and pSB1A3 had purified with mycrocon, and 1-23L had extracted from a gel before.

electrophoresed 1ul of 1-14K and 1-23L added 0.5ul of 6×sample buffer to estimate concentration of each solution.
estimated concentration from photo of electrophoresys.But I forgot to electrophorese pSB1A3 solution with the other samples,so pSB1A3 solution was done by other person.
made digestion recipes based on each concentrations(below). Why pSB1A3 recipe is two, because I firstly made 30ul of pSB1A3 solution, but I found it was insufficient to ligate parts, so I made more 50ul of it after.

put each solutions into 37℃ incubator.1-23L solution was put about a half and two hours,1-14K solution was done about a half and an hour,30ul of pSB1A3 solution was done about an hour,and 50ul of pSB1A3 was done about a half hour.

electrophoresed each solutions added 6×sample buffer.put 12uls each into wells of a gel.

cannot see 1-23L because of overflowing.extracted the other samples from a gel.dissolve them with 50ul of Nuclease free water.And they were stocked to freeze in -20℃.

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-# put each solutions into 37℃ incubator.1-23L solution was put about a half and two hours,1-14K solution was done about a half and an hour,30ul of pSB1A3 solution was done about an hour,and 50ul of pSB1A3 was done about a half hour.
+* put each solutions into 37℃ incubator.1-23L solution was put about a half and two hours,1-14K solution was done about a half and an hour,30ul of pSB1A3 solution was done about an hour,and 50ul of pSB1A3 was done about a half hour.
-# electrophoresed each solutions added 6×sample buffer.put 12uls each into wells of a gel.
+* electrophoresed each solutions added 6×sample buffer.put 12uls each into wells of a gel.
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