Week of 6/21

==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



<center>[[Image:6-22-10.jpg]]</center>



*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


<center>[[Image:6-22-10 001.jpg]]</center>

*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


<center>[[Image:6-24-10.jpg]]</center>


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



<center>[[Image:6-25-10.jpg]]</center>




*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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