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From 2010.igem.org

Contents 1 June 21st 2 June 22nd 3 June 23rd 4 June 24th 5 June 25th

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

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