"
Week of 7/26
From 2010.igem.org
(Difference between revisions)
| Line 26: | Line 26: | ||
*Digests: | *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) | ||
<center>[[Image:7-27-10.jpg]]</center> | <center>[[Image:7-27-10.jpg]]</center> | ||
| + | *A second gel was ran from the digests that were done today. The gel order is provided below: | ||
| + | -Ladder | ||
| + | -pSB1T3 yellow | ||
| + | -pJ241 | ||
| + | -A-E | ||
<center>[[Image:7-27-10 001.jpg]]</center> | <center>[[Image:7-27-10 001.jpg]]</center> | ||
| + | *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 | ||
<center>[[Image:7-28-10.jpg]]</center> | <center>[[Image:7-28-10.jpg]]</center> | ||
| + | *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 | ||
<center>[[Image:7-29-10.jpg]]</center> | <center>[[Image:7-29-10.jpg]]</center> | ||
| + | -All bands showed up correctly and so all three were cut out and then gel purified. | ||
| + | |||
| + | |||
| + | ==July 30th== | ||
[https://2010.igem.org/Team:Penn_State/Notebook Return to Notebook] | [https://2010.igem.org/Team:Penn_State/Notebook Return to Notebook] | ||
Revision as of 18:25, 30 August 2010
Contents |
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.
