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Week of 6/28
From 2010.igem.org
(Difference between revisions)
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(→June 30) |
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| - | *A second gel was | + | *A second gel was run the same day. The order is provided below: |
- Ladder | - Ladder | ||
- B1002 | - B1002 | ||
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- K274100 | - K274100 | ||
- K112022 | - K112022 | ||
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==July 1== | ==July 1== | ||
Revision as of 14:53, 5 August 2010
Contents |
June 28
- Retransform:
-R0061 -R0062 -R0063 -K091146
- Inventory
-Good Sequencing:
o Backbone - C3RFP
o pLux - K091146
o Lysis - K112022
o pLux - R0063
o pLux - R1062
o CI+RBS - K081007
o Cons.Prom- J23116
o Cons.Prom- J23108
o Pigment - K274100
o LuxI - C0261
o Lysis - K112808
o pLac - R0010
o LacI+RBS - J24679
- To grow:
- K091146 - R0063 - J23108 - J23116
- Electrocompetent cells were made.
June 29
Gel Order:
- J23108 - J23108 (Digest 2) - R0063 - R0063 (Digest 2) - K081007 - K081007(Digest 2) - J23116 - J23116 (Digest 2)
- Gel extract was done using Digest number 1. In this gel we ran the same part twice but from two different digests. Digest 1 was a 6 hour digest in the 37 degree Celsius water bath while Digest 2 was a 1 hour digest in the same water bath. As you can see, the results do not differ all that noticeably. That being established we went back to doing 1 hour digests.
- The bands that were cut out are provided below:
- J23108 - R0063 - J23116 - K081007
All these bands were purified.
June 30
- On this gel, we ran the Colony PCR and the digests. The order is as follows:
Colony PCR:
- Ladder - A-J
Digests:
- Ladder - K091107 (digest 2) - K091107 - K091146 (digest 2) - K091146 - J23116+J37033 (digest 2) - J23116+J37033 - R0010 (digest 2) - R0010
- The bands that were cut are marked with a square around them in the picture above. They were also then purified.
- 3 bands that were cut:
- K091107 - K091146 - R0010
- A second gel was run the same day. The order is provided below:
- Ladder - B1002 - Lux R 2 - C0261+J24679
- As you can see, the only 2 bands that were cut were B1002 and C0261+J24679
- Grow for tomorrow:
- K274100 - K112022
July 1
- Flasks minipreped:
- B1006 - K274100 - J23100
- Digests:
- B1006 (EX) - K274100 (XP) - J23100 (SP)
- Sent to the sequencing center:
1) B1002 2) B1006 3) S01414 4) J23100 5) C0261+J24679
Gel order:
- Ladder - B1006 - K274100 - J23100
- All the bands appeared where we wanted them to so we cut out all three bands and then purified them.
- Ligations:
Vector Inserts
1)J23100 K274100 2)J23100 S01414 3)R0063 a)K274100 4)K091146 b)C0261+J24679 5)K091107 6)R0062 7)R0010 K081007
- Tomorrow:
- Develop primers to figure out if K274100 and K112808 are correct.
July 2
- Colony PCR's ran on gel.The order is as follows:
- Ladder - 1A, 1B, 1C - 2A, 2B, 2C - 3A, 3B, 3C - Ladder - 4A, 4B, 4C - 5A, 5B, 5C - 6A, 6B, 6C - 7A, 7B, 7C
Unfortunately, we accidentally put ladder in all the Colony products. The picture is provided above.
