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Tuesday, July 26
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So I used a protocol suggested by a colleague here in TU's molecular biology group and put the spin down products from tubes labeled P1, T1, T2, and R2 in a 20% solution of sterile glycerol, in special cryo-tubes that the same colleague donated, and placed these in the -80°C freezer in the bottom shelf of the freezer in the autoclave room across the hall from Rm 491. | So I used a protocol suggested by a colleague here in TU's molecular biology group and put the spin down products from tubes labeled P1, T1, T2, and R2 in a 20% solution of sterile glycerol, in special cryo-tubes that the same colleague donated, and placed these in the -80°C freezer in the bottom shelf of the freezer in the autoclave room across the hall from Rm 491. | ||
| - | Spun the other tubes more gently for a dokee period of time, so probably (almost certainly) sacrificed some cells and DNA, added | + | Spun the other tubes more gently for a dokee period of time, so probably (almost certainly) sacrificed some cells and DNA, added 50µl Tris pH 8.0 and placed in freezer in 491 for cleanup at a later date. |
Revision as of 23:56, 28 July 2011
Dr Liz's comments:
Lab opened about 1:20pm. Only person present was Dr Liz.
Trying to make good sense out it5 what is in the freezer. Our single-colony clones were in the cold room, still in the original tubes in which they were incubated. I decided to spin them down to get smaller volumes with the DNA. I should have known that some tubes cannot withstand 12000rpm. Lost two tubes that collapsed in the centrifuge. The lost samples were labeled P1 and R2. Notice this means one out of the four "P" colonies was lost and one of the four "R" colonies was lost.
Decided to take precautions to preserve some of each of the three types (P, R, T) of transformed E. coli colonies (We suppose and hope they are transformed, as they did survive amp-laced LB agar).
So I used a protocol suggested by a colleague here in TU's molecular biology group and put the spin down products from tubes labeled P1, T1, T2, and R2 in a 20% solution of sterile glycerol, in special cryo-tubes that the same colleague donated, and placed these in the -80°C freezer in the bottom shelf of the freezer in the autoclave room across the hall from Rm 491.
Spun the other tubes more gently for a dokee period of time, so probably (almost certainly) sacrificed some cells and DNA, added 50µl Tris pH 8.0 and placed in freezer in 491 for cleanup at a later date.
