Production/Competent Cells
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 | 3 | 0 | 0 | 4 | Yes |
| 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 | 7 | 0 | 0 | 4 | Yes |
| 004 | NEB 10b (002) | KN+VS | 9/18/11 | 20C, 240rpm | 18 | 500ml | 0.13 | NA | NA | NA | NA | NA | NA | NA | NA |
| 005 | NEB 10b (002) | KN+VS | 9/18/11 | 20C, 240rpm | 18 | 500ml | 0.14 | NA | NA | NA | NA | NA | NA | NA | NA |
| 006 | NEB 10b (002) | KN+VS | 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 | Yes |
| 007 | NEB 10b (002) | KN+VS | 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 | Yes |
| 008 | NEB 10b (002) | KN+VS | 9/22/11 | 20C, 240rpm | 21 | 500ml | 0.27 | 0.68 | TBD | TBD | 8 | 0 | 0 | 4 | Yes |
| 009 | NEB 10b (002) | KN+VS | 9/22/11 | 20C, 240rpm | 22 | 500ml | 0.16 | NA | NA | NA | NA | NA | NA | NA | 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
First comp cell batch to use seed stock batch #002. 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). Test transformations were performed with the first and last 250ul aliquot.
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). Test transformations were performed with the first and last 250ul aliquot.
008
Cells grew slowly; temperature was increased to 23C at 19 hours. Cells were OD'ed multiple times: 18 hours (0.09), 19 hours (0.11), and 21 hours (0.27). Final ice incubation step was approximately 25 minutes.
009
Cells grew very slowly; temperature was increased to 23C at 19 hours. Cells were OD'ed multiple tiems: 18 hours (0.04), 19 hours (0.06), 21 hours (0.13), and 22 hours (0.16). Batch was discarded.