Difference between revisions of "Production/Competent Cells"

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<tr><td>004 <td>NEB 10b (002)<td> KN+VS <td>9/18/11 <td>20C, 240rpm <td> 18 <td>500ml <td>0.13<td> NA <td> NA <td> NA <td> NA <td> NA <td> NA <td> NA <td> NA </tr>
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<tr style="background-color:#E0E0E0"><td>004 <td>NEB 10b (002)<td> KN+VS <td>9/18/11 <td>20C, 240rpm <td> 18 <td>500ml <td>0.13<td> NA <td> NA <td> NA <td> NA <td> NA <td> NA <td> NA <td> NA </tr>
 
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<tr><td>005 <td>NEB 10b (002)<td> KN+VS <td>9/18/11 <td>20C, 240rpm <td> 18 <td>500ml <td>0.14<td> NA <td> NA <td> NA <td> NA <td> NA <td> NA <td> NA <td> NA </tr>
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<tr style="background-color:#E0E0E0"><td>005 <td>NEB 10b (002)<td> KN+VS <td>9/18/11 <td>20C, 240rpm <td> 18 <td>500ml <td>0.14<td> NA <td> NA <td> NA <td> NA <td> NA <td> NA <td> NA <td> NA </tr>
 
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<tr><td> 009 <td>NEB 10b (002)<td> KN+VS <td>9/22/11 <td>20C, 240rpm <td> 22 <td>500ml <td>0.16<td> NA <td> NA <td> NA <td> NA <td> NA <td> NA <td> NA <td> NA </tr>
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<tr style="background-color:#E0E0E0"><td> 009 <td>NEB 10b (002)<td> KN+VS <td>9/22/11 <td>20C, 240rpm <td> 22 <td>500ml <td>0.16<td> NA <td> NA <td> NA <td> NA <td> NA <td> NA <td> NA <td> NA </tr>
 
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<tr><td> 010 <td>NEB 10b (002)<td> VS+KN <td>9/27/11 <td>20C, 240rpm <td> 22 <td>500ml <td>0.38<td>1.52<td>88, 74, 81, 40, 47, 62 (65.3) <td> 9.800*10^7 <td> 2 <td> 0 <td> 0<td> 5 <td> Yes </tr>
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<tr><td> 011 <td>NEB 10b (002)<td> KN+VS <td>9/27/11 <td>20C, 240rpm <td> 22 <td>500ml <td>0.46<td>1.47<td> 24, 41, 48, 20, 22, 32 (31.2) <td> 4.675*10^7 <td>8<td> 0 <td> 0 <td> 4 <td> Yes </tr>
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===009===
 
===009===
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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.
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Cells grew very slowly; temperature was increased to 23C at 19 hours. Cells were OD'ed multiple times: 18 hours (0.04), 19 hours (0.06), 21 hours (0.13), and 22 hours (0.16). Batch was discarded.
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===010===
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Minimal dry ice available. "Last" 250ul aliquot was aliquoted after finishing 4 plates.
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===011===
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Minimal dry ice available. "Last" 250ul aliquot was second to last 250ul aliquot.

Latest revision as of 20:20, 20 October 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 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 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
010 NEB 10b (002) VS+KN 9/27/11 20C, 240rpm 22 500ml 0.381.5288, 74, 81, 40, 47, 62 (65.3) 9.800*10^7 2 0 0 5 Yes
011 NEB 10b (002) KN+VS 9/27/11 20C, 240rpm 22 500ml 0.461.47 24, 41, 48, 20, 22, 32 (31.2) 4.675*10^7 8 0 0 4 Yes

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 times: 18 hours (0.04), 19 hours (0.06), 21 hours (0.13), and 22 hours (0.16). Batch was discarded.

010

Minimal dry ice available. "Last" 250ul aliquot was aliquoted after finishing 4 plates.

011

Minimal dry ice available. "Last" 250ul aliquot was second to last 250ul aliquot.