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From 2010.igem.org

Contents 1 July 26th 2 July 27th 3 July 28th 4 July 29th 5 July 30th 6 August 1st

July 26th

• Colony PCR:

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

• Digests:

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

• Ligations:

 -Lux+RBS+A3

• Gel Order:

 -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.

July 27th

• Digests:

 - 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

• Items sent to sequencing:

• Sent to sequencing:

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

• Grow for tomorrow:

-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.

July 28th

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.

• We also grew up pSB1T3 red, pSB1T3 yellow, and pJ241

July 29th

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

• Digests:

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

• Grow for tomorrow:

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

• Minipreps done today:

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

• Sent to sequencing:

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

• Gel:

-Ladder
-pJ241
-pSB1T3 red
-pSB1T3 yellow

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

July 30th

• Minipreps done today:

-K145150
-R0063
-K3
-RFP+RBS

• Digests:

-Lux+RBS+A3 (XP)
-J23100 (ES+phosphatase)
-J24679+B1002 (XP)
-C0261 (EP+phosphatase)
-RFP+RBS (XP)
-R0062 (EP+phosphatase)
-R0063 (EP+phosphatase)
-K145150 (EP+phosphatase)
-K091107 (EP+phosphatase)
-K091146 (EP+phosphatase)

• Ligations:

- Lux+RBS+A3 + J23100 + C3
- J24679+B1002 + C0261 + C3
- RFP+RBS + R0062 + C3
- RFP+RBS + R0063 + C3
- RFP+RBS + K145150 + C3
- RFP+RBS + K091107 + C3
- RFP+RBS + K091146 + C3

• Grow for next week:

-R0061
-Electrocompetent cells

August 1st

• Colony PCR

1) J23100+Lux+RBS+C3
2) R0062+RFP+RBS+C3 redo
3) K091107+RFP+RBS+C3 redo
4) K091146+RFP+RBS+C3 redo

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