Difference between revisions of "Production/Competent Cells"
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| − | <tr><td> 006 <td>NEB 10b (002)<td> KN <td>9/19/11 <td>20C, 240rpm <td> | + | <tr><td> 006 <td>NEB 10b (002)<td> KN <td>9/19/11 <td>20C, 240rpm <td> 21 <td>500ml <td>0.28<td> 0.74 <td> 20, 29, 17, 10, 10, 12 (16.3) <td> 2.45*10^7 <td> 4 <td> 0 <td> 0 <td> 4 <td> NA </tr> |
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Revision as of 18:03, 23 September 2011
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 | Checklist pdf |
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| 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 | NA |
| 002 | NEB 10b | VS+KN | 9/12/11 | 20C, 240rpm | 16 | 500ml | 0.40 | 1.12 | 91, 140, 131, 45, 66, 30 (83.3) | 1.258*10^8 | ?? | 0 | 0 | 4 | ?? |
| 003 | NEB 10b | VS+KN | 9/12/11 | 20C, 240rpm | 16 | 500ml | 0.23 | 0.71 | 127, 145, 72, 83, 93, 60 (96.7) | 1.450*10^8 | ?? | 0 | 0 | 4 | ?? |
| 004 | NEB 10b (002) | KN | 9/18/11 | 20C, 240rpm | 18 | 500ml | 0.13 | NA | NA | NA | NA | NA | NA | NA | NA |
| 005 | NEB 10b (002) | KN | 9/18/11 | 20C, 240rpm | 18 | 500ml | 0.14 | NA | NA | NA | NA | NA | NA | NA | NA |
| 006 | NEB 10b (002) | KN | 9/19/11 | 20C, 240rpm | 21 | 500ml | 0.28 | 0.74 | 20, 29, 17, 10, 10, 12 (16.3) | 2.45*10^7 | 4 | 0 | 0 | 4 | NA |
| 007 | NEB 10b (002) | KN | 9/19/11 | 20C, 240rpm | 20 | 500ml | 0.23 | 0.76 | 14, 10, 15, 9, 9, 13 (11.6) | 1.750*10^7 | 7 | 0 | 0 | 4 | NA |
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.
Notes: While ODing culture, cells were not kept on ice/stored in fridge. Poured into centrifuge tubes on ice, but tubes were not pre-chilled.
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.
Notes: While ODing culture, cells were not kept on ice/stored in fridge. Poured into centrifuge tubes on ice, but tubes were not pre-chilled.
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.
Notes: While ODing culture, cells were not kept on ice/stored in fridge. Poured into centrifuge tubes on ice, but tubes were not pre-chilled.
004
Cells grew very slowly; low OD even after 18 hours of culture. Batch was discarded.
005
Cells grew very slowly; low OD even after 18 hours of culture. Batch was discarded.
006
Media was inoculated at 4pm on Day 1 to allow for longer growth period. Cells grown in 1L flask. Cells were OD'ed multiple times: 16 hours (0.12), 16.5 hours (0.12), 18.5 hours (0.15), and 21 hours (0.28).
007
Media was inoculated at 4pm on Day 1 to allow for longer growth period. Cells grown in 1L flask. Cells were OD'ed multiple times: 16 hours (0.13), 16.5 hours (0.14), 18.5 hours (0.22), and 20 hours (0.23).