Week of 6/21

From 2010.igem.org

(Difference between revisions)
(June 22nd)
(June 23rd)
Line 97: Line 97:
     - K274100
     - K274100
-
*Digests:
+
*[https://2010.igem.org/DNA_Digest Digests]:
     - K091107 (SP)
     - K091107 (SP)
     - R0010  (SP)
     - R0010  (SP)

Revision as of 15:26, 5 August 2010

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


6-22-10.jpg


  • 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


6-22-10 001.jpg
  • 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
   - 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


6-24-10.jpg


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


6-25-10.jpg



  • 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




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