Production/Competent Cells

Revision as of 18:03, 23 September 2011 by Kitwa (Talk | contribs) (Batch Notes)


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 Values20C160.3 or lower 1 - 1.5100 - 4005*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 20 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).