Week of 6/21
From 2010.igem.org
Contents |
June 21st
- During this week we began doing 6 hour digests at the recommendation of Dr. Salis to cut down on background
Sent to the sequencing center:
Base Pairs 1)C3RFP 681 2)K091146 1266 3)K112022 1499 4)K145150 66 5)R0063 151 6)R1062 56 7)K081007 796
- We made 1 liter of LB media.
June 22nd
- Flasks Minipreped:
- J23108 - J23116 - J37033 - K091107 - C0261 - K274100 - C3RFP - K112808
- All the minipreps were digested with the following enzymes:
- J23108 (SP) - J23116 (SP) - J37033 (XP + phosphatase) - K091107(SP) - C0261 (XP + phosphatase) - K274100(XP + phosphatase) - C3RFP (EX) - K112808(ES + phosphatase)
- We also did a PCR on
-1. K081007 + R0010 -2. J23116 + J37033 -3. J23108 + J37033
- 3b and 3c did not turn out for us as planned.
- Ligations:
- J23108 + J37033 - J23116 + J37033 - K091107 + C0261 - K091107 + K274100 - C3RFP + K112808
- Transformations:
- B0010 Plate 1 Well 13D - R0010 Plate 1 Well 1D - K081007 Plate 2 Well 10J - J24679 Plate 1 Well 7L
- Gel order:
- Ladder - J23116 - J23108 - K091107 - C3RFP
- This gel turned out well for us, C3RFP was visible but faint.
June 23rd
- Flasks mini prepped:
- K091107 - R0010 - J24679 - J23108 - J23116 - C0261 - K081107 - B0010 - J37033 - K274100
- Digests:
- K091107 (SP) - R0010 (SP) - J24679 (ES) - J23108 (SP) - J23116 (SP) - CO261 (XP) - K081107 (XP) - B0010 (EX) - J37033 (XP) - K274100 (XP)
- Sent to sequencing center:
1) R0010 2) B0010 3) K081107 4) J24679
- Transform today:
- R0061 Plate 1 Well 14P - R0062 Plate 1 Well 6O - R0063 Plate 1 Well 8A - K091146 Plate 2 Well 8H
June 24th
Gel Order:
- Ladder - J24679 - K091107 - J37033 - J23108 - K274100 - B0010 - R0010 - K081107 - CO261 - J23116
RE-DIGEST:
- CO261 - K081107
RE-Transform:
- J37033 - K091107
- Sent to sequencing:
1) R0061 2) R0062 3) R0063 4) K091146
June 25th
- Sent to sequencing:
1) K091107 2) J37033
- Digests:
- K091107 (SP) - J23108 (SP) - J23116 (SP) - CO261 (XP) - R0010 (SP) - K274100 (XP) - J37033 (XP) - J24679 (ES) - K081007 (XP)
The 4 that were sent to get sequenced yesterday were ran on a gel. The gel order is provided below:
- Ladder - K091146 - R0061 - R0062 - R0063
- This gel was perfect for us. We were able to see what we needed to see and extract the correct pieces.
- Ligations:
- K091107 + K274100 - J23108 + J37033 - J23116 + J37033 - C0261 + J24679 - R0010 + K081007