Week of 6/21

From 2010.igem.org

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-
==June 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:
Sent to the sequencing center:
Line 11: Line 15:
   7)K081007    796
   7)K081007    796
-
*We made 1 liter of LB media.  
+
*We made 1 liter of LB media.
-
 
+
==June 22nd==
-
==June 22==
+
*Flasks Minipreped:
*Flasks Minipreped:
-
  -J23108
+
    - J23108
-
  -J23116
+
    - J23116
-
  -J37033
+
    - J37033
-
  -K091107
+
    - K091107
-
  -C0261
+
    - C0261
-
  -K274100
+
    - K274100
-
  -C3RFP
+
    - C3RFP
-
  -K112808
+
    - K112808
-
*All the minipreps were cut with the following enzymes:
+
*All the minipreps were digested with the following enzymes:
-
  -J23108 (SP)
+
    - J23108 (SP)
-
  -J23116 (SP)
+
    - J23116 (SP)
-
  -J37033 (XP + phosphatase)
+
    - J37033 (XP + phosphatase)
-
  -K091107(SP)
+
    - K091107(SP)
-
  -C0261  (XP + phosphatase)
+
    - C0261  (XP + phosphatase)
-
  -K274100(XP + phosphatase)
+
    - K274100(XP + phosphatase)
-
  -C3RFP  (EX)
+
    - C3RFP  (EX)
-
  -K112808(ES + phosphatase)
+
    - K112808(ES + phosphatase)
-
*Ligations:
 
-
- J23108 + J37033
 
-
- J23116 + J37033
 
-
- K091107 + C0261
 
-
- K091107 + K274100
 
-
- C3RFP + K112808
 
-
*Transformations:
+
*We also did a PCR on
-
- B0010    Plate 1 Well 13D
+
-
- R0010    Plate 1 Well 1D
+
-
- K081007  Plate 2 Well 10J
+
-
- J24679  Plate 1 Well 7L
+
 +
    -1. K081007 + R0010
 +
    -2. J23116 + J37033
 +
    -3. J23108 + J37033
 +
<center>[[Image:6-22-10.jpg]]</center>
-
*We also did a PCR on K081007 + R0010, J23116 + J37033, and 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>
<center>[[Image:6-22-10 001.jpg]]</center>
 +
*This gel turned out well for us, C3RFP was visible but faint.
 +
==June 23rd==
-
==June 23==
 
-
*Transform today:  
+
*Flasks mini prepped:
-
  - R0061  Plate 1 Well 14P
+
    - K091107
-
  - R0062  Plate 1 Well 6O
+
    - R0010
-
  - R0063  Plate 1 Well 8A
+
    - J24679
-
  - K091146 Plate 2 Well 8H
+
    - J23108
 +
    - J23116
 +
    - C0261
 +
    - K081107
 +
    - B0010
 +
    - J37033
 +
    - K274100
*Digests:
*Digests:
-
  - K091107 (SP)
+
    - K091107 (SP)
-
  - R0010  (SP)
+
    - R0010  (SP)
-
  - J24679  (ES)
+
    - J24679  (ES)
-
  - J23108  (SP)
+
    - J23108  (SP)
-
  - J23116  (SP)
+
    - J23116  (SP)
-
  - CO261  (XP)
+
    - CO261  (XP)
-
  - K081107 (XP)
+
    - K081107 (XP)
-
  - B0010  (EX)
+
    - B0010  (EX)
-
  - J37033  (XP)
+
    - J37033  (XP)
-
  - K274100 (XP)
+
    - K274100 (XP)
-
*Sent to get sequenced:
+
*Sent to sequencing center:
-
  1) R0010
+
    1) R0010
-
  2) B0010
+
    2) B0010
-
  3) K081107
+
    3) K081107
-
  4) J24679
+
    4) J24679
-
==June 24==
+
*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:
Gel Order:
-
  1) Ladder
+
    - Ladder
-
  2) J24679
+
    - J24679
-
  3) K091107
+
    - K091107
-
  4) J37033
+
    - J37033
-
  5) J23108
+
    - J23108
-
  6) K274100
+
    - K274100
-
  7) B0010
+
    - B0010
-
  8) R0010
+
    - R0010
-
  9) K081107
+
    - K081107
-
  10)CO261
+
    - CO261
-
  11)J23116
+
    - J23116
Line 113: Line 142:
RE-DIGEST:
RE-DIGEST:
-
- CO261
+
    - CO261
-
- K081107
+
    - K081107
RE-Transform:
RE-Transform:
-
- J37033
+
    - J37033
-
- K091107
+
    - K091107
*Sent to sequencing:
*Sent to sequencing:
-
1) R0061
+
    1) R0061
-
2) R0062
+
    2) R0062
-
3) R0063
+
    3) R0063
-
4) K091146
+
    4) K091146
-
==June 25==
+
==June 25th==
*Sent to sequencing:
*Sent to sequencing:
-
1) K091107
+
    1) K091107
-
2) J37033
+
    2) J37033
*Digests:
*Digests:
-
  - K091107 (SP)
+
    - K091107 (SP)
-
  - J23108  (SP)
+
    - J23108  (SP)
-
  - J23116  (SP)
+
    - J23116  (SP)
-
  - CO261  (XP)
+
    - CO261  (XP)
-
  - R0010  (SP)
+
    - R0010  (SP)
-
  - K274100 (XP)
+
    - K274100 (XP)
-
  - J37033  (XP)
+
    - J37033  (XP)
-
  - J24679  (ES)
+
    - J24679  (ES)
-
  - K081007 (XP)
+
    - K081007 (XP)
 +
 
-
*Ligations:
 
-
  - K091107 + K274100
 
-
  - J23108  + J37033
 
-
  - J23116  + J37033
 
-
  - C0261  + J24679
 
-
  - R0010  + K081007
 
The 4 that were sent to get sequenced yesterday were ran on a gel. The gel order is provided below:
The 4 that were sent to get sequenced yesterday were ran on a gel. The gel order is provided below:
-
- Ladder
+
    - Ladder
-
- K091146
+
    - K091146
-
- R0061
+
    - R0061
-
- R0062
+
    - R0062
-
- R0063
+
    - R0063
 +
 
 +
 
<center>[[Image:6-25-10.jpg]]</center>
<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
 +
 +
 +
 +
 +
 +
 +
 +
[https://2010.igem.org/Team:Penn_State/Notebook Return to Notebook]

Latest revision as of 15:41, 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
  • 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


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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