Week of 7/26

Revision as of 18:25, 30 August 2010 by Anisha (Talk | contribs)

July 26th

 -J24679+B1002+K3
 -J24679+B1006+K3
 -Lux+RBS+B1006+K3 redo
 -Lux+RBS+B1002+K3

 -Lux+RBS (EP)
 -A3 (EP+Phosphatase)

 -Lux+RBS+A3

 -Ladder
 -J24679+B1002+K3 (A-E)
 -J24679+B1006+K3 (A-E)

The bands for 1B and 1D showed up so they were grown up accordingly.

 - C0261 (ES)
 - R0010 (ES)
 - J23100(ES)
 - J24679(XP)
 - P0451 (XP)
 - LuxR+RBS (XP)

The colony PCR that was done yesterday was ran on a gel. The gel order is as follows:

- Ladder
- J24679+B1006+K3 (E)
- Lux+RBS+B1006+K3 redo (A-E)
- Lux+RBS+B1002+K3 (A-E)

A second gel was ran from the digests that were done today. The gel order is provided below:

-Ladder
-pSB1T3 yellow
-pJ241
-A-E

1) J24679+B1002 B (Reverse)
2) J24679+B1002 D (Reverse)
3) LuxR+RBS+A3 C (Forward+Reverse)
4) LuxR+RBS+A3 A (Forward+Reverse)

-C0261
-J23100
-pJ241
-pSB1T3 yellow

All 3 pigments were streaked onto the plate to make new stocks.

Only pJ241 was ligated because there was a top and bottom band and we were not sure which was the actual pigment we extracted.

Colony PCR Gel:

- Ladder
-13A-E
-14A-D
 13: P0451+R0010
 14: J23100+LuxR+RBS

Since 13B and C grew, we sent those to sequencing with forward and reverse primers. Lux+RBS+A3 was also sent to sequencing with the same primers.

J24679+B1002 (B) and (D) were resent to the sequencing center using the R primer.

- R0061(ES)
- R0062(ES)
- R0063(ES)
- K091146(ES) 
- K091107(ES)
- pJ241 (XP)
- pSB1T3 yellow (XP)
- K3 (EP)
- T3 (EP)

- K145150
- K3 
- R0063
- R0061
- RFP+RBS

-P0451+R0010 B
-R0451+R0010 C
-pSBT13 yellow
-pJ241
-pSBT13 red

1)pSBT13 red (forward+reverse)
2)pSBT13 yellow (forward+reverse)

-Ladder
-pJ241
-pSB1T3 red
-pSB1T3 yellow

-All bands showed up correctly and so all three were cut out and then gel purified.