Difference between revisions of "Production/Competent Cells"
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Revision as of 15:10, 14 September 2011
Contents
Batch Summary
Use this page to keep track of competent cell production. See the competent cell protocol for more information.
| Batch | Strain | Maker | Date Started | Temp, RPM | Incubation (hr) | Culture Volume | Culture OD (600nm) | Competent OD (600nm) | Cell Count (AVG) | Efficiency | 250ul Aliquots | 500ul Aliquots | 1ml Aliquots | 96w Plate |
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Expected Values | 20C | 16 | 0.3 or lower | 1 - 1.5 | 100 - 400 | 5*10^8 - 5*10^9 | ||||||||
| 001 | NEB 10b | VS+KN | 8/17/11 | 21C, 250rpm | 16 | 500ml | 0.4 | 1.35 | 67,88,112 (89) | 1.335*10^8 | 20 | 20 | 10 | 0 |
| 002 | NEB 10b | VS+KN | 9/12/11 | 20C, 250rpm | 16 | 500ml | 0.40 | 1.12 | ??, ??, ?? (??) | ?? | ?? | 0 | 0 | 4 |
| 003 | NEB 10b | VS+KN | 9/12/11 | 20C, 250rpm | 16 | 500ml | 0.23 | 0.71 | ??, ??, ?? (??) | ?? | ?? | 0 | 0 | 4 |
Batch Notes
001
This is the first batch attempting to fully standardize competent cell production. Also Kitwa's first batch! And a resounding success!
The efficiency test/test transformation used a single 250ul aliquot (randomly chosen). Three transformations were done using our standard template and protocol. Important: In following productions we will test the first and last of the aliquots.
002
OD600nm value at 15.5 hours was 0.26, and cells were re-placed in the shaker for another 0.5 hours. OD value afterward was 0.40.
003
OD600nm value at 15.5 hours was 0.16, and cells were re-placed in the shaker for another 0.5 hours. OD value afterward was 0.23.