Week of 6/28

From 2010.igem.org

(Difference between revisions)
Line 1: Line 1:
 +
==June 28==
 +
 +
* Retransform:
 +
  -R0061
 +
  -R0062
 +
  -R0063
 +
  -K091146
 +
 +
*Inventory
 +
-Good Sequencing:
 +
    o Backbone - C3RFP
 +
    o pLux    - K091146
 +
    o Lysis    - K112022
 +
    o pLux    - R0063
 +
    o pLux    - R1062
 +
    o CI+RBS  - K081007
 +
    o Cons.Prom- J23116
 +
    o Cons.Prom- J23108
 +
    o Pigment  - K274100
 +
    o LuxI    - C0261
 +
    o Lysis    - K112808
 +
    o pLac    - R0010
 +
    o LacI+RBS - J24679
 +
 +
*To grow:
 +
    - K091146
 +
    - R0063
 +
    - J23108
 +
    - J23116
 +
 +
*Electrocompetent cells were made.
 +
 +
 +
==June 29==
 +
 +
Gel Order:
 +
    - J23108
 +
    - J23108 (Digest 2)
 +
    - R0063 
 +
    - R0063  (Digest 2)
 +
    - K081007
 +
    - K081007(Digest 2)
 +
    - J23116
 +
    - J23116 (Digest 2)
 +
 +
<center>[[Image:6-29-10.jpg]]</center>
<center>[[Image:6-29-10.jpg]]</center>
 +
*Gel extract was done using Digest number 1.
 +
 +
*The bands that were cut out are provided below:
 +
    - J23108
 +
    - R0063
 +
    - J23116
 +
    - K081007
 +
 +
All these bands were purified.
 +
 +
 +
==June 30==
 +
 +
 +
*On this gel, we ran the Colony PCR and the digests. The order is as follows:
 +
 +
Colony PCR:
 +
  - Ladder
 +
  - A-J
 +
 +
Digests:
 +
  - Ladder
 +
  - K091107 (digest 2)
 +
  - K091107
 +
  - K091146 (digest 2)
 +
  - K091146
 +
  - J23116+J37033 (digest 2)
 +
  - J23116+J37033
 +
  - R0010 (digest 2)
 +
  - R0010
<center>[[Image:6-30-10.jpg]]</center>
<center>[[Image:6-30-10.jpg]]</center>
 +
*The bands that were cut are marked with a square around them in the picture above. They were also then purified.
 +
* 3 bands that were cut:
 +
    - K091107
 +
    - K091146
 +
    - R0010
 +
 +
 +
 +
*A second gel was ran the same day. The order is provided below:
 +
  - Ladder
 +
  - B1002
 +
  - Lux R 2
 +
  - C0261+J24679
<center>[[Image:6-30-10 001.jpg]]</center>
<center>[[Image:6-30-10 001.jpg]]</center>
 +
*As you can see, the only 2 bands that were cut were B1002 and C0261+J24679
 +
 +
 +
*Grow for tomorrow:
 +
  - K274100
 +
  - K112022
 +
 +
 +
 +
==July 1==
 +
 +
 +
*Flasks minipreped:
 +
    - B1006
 +
    - K274100
 +
    - J23100
 +
 +
*Digests:
 +
    - B1006 (EX)
 +
    - K274100 (XP)
 +
    - J23100 (SP)
 +
 +
*Sent to the sequencing center:
 +
  1) B1002
 +
  2) B1006
 +
  3) S01414
 +
  4) J23100
 +
  5) C0261+J24679
 +
 +
 +
Gel order:
 +
  - Ladder
 +
  - B1006
 +
  - K274100
 +
  - J23100
<center>[[Image:7-1-10.jpg]]</center>
<center>[[Image:7-1-10.jpg]]</center>
 +
*All the bands appeared where we wanted them to so we cut out all three bands and then purified them.
 +
 +
 +
 +
*Ligations:
 +
  Vector        Inserts
 +
1)J23100        K274100
 +
2)J23100        S01414
 +
3)R0063          a)K274100
 +
4)K091146        b)C0261+J24679
 +
5)K091107
 +
6)R0062
 +
7)R0010          K081007
 +
 +
*Tomorrow:
 +
- Develop primers to figure out if K274100 and K112808 are correct.
 +
 +
 +
 +
==July 2==
 +
 +
 +
*Colony PCR's ran on gel.The order is as follows:
 +
    - Ladder
 +
    - 1A, 1B, 1C
 +
    - 2A, 2B, 2C
 +
    - 3A, 3B, 3C
 +
 +
    - Ladder
 +
    - 4A, 4B, 4C
 +
    - 5A, 5B, 5C
 +
    - 6A, 6B, 6C
 +
    - 7A, 7B, 7C
<center>[[Image:7-2-10.jpg]]</center>
<center>[[Image:7-2-10.jpg]]</center>
 +
 +
Unfortunately, we accidentally put ladder in all the Colony products. The picture is provided above.

Revision as of 18:05, 4 August 2010

Contents

June 28

  • Retransform: 
  -R0061
  -R0062
  -R0063
  -K091146
  • Inventory 
-Good Sequencing:
    o Backbone - C3RFP
    o pLux     - K091146
    o Lysis    - K112022
    o pLux     - R0063
    o pLux     - R1062
    o CI+RBS   - K081007
    o Cons.Prom- J23116
    o Cons.Prom- J23108
    o Pigment  - K274100
    o LuxI     - C0261
    o Lysis    - K112808
    o pLac     - R0010
    o LacI+RBS - J24679
  • To grow: 
   - K091146
   - R0063
   - J23108
   - J23116
  • Electrocompetent cells were made.


June 29

Gel Order: 

   - J23108
   - J23108 (Digest 2)
   - R0063  
   - R0063  (Digest 2)
   - K081007
   - K081007(Digest 2)
   - J23116 
   - J23116 (Digest 2)


6-29-10.jpg
  • Gel extract was done using Digest number 1.
  • The bands that were cut out are provided below: 
   - J23108
   - R0063
   - J23116
   - K081007

All these bands were purified.


June 30

  • On this gel, we ran the Colony PCR and the digests. The order is as follows:

Colony PCR: 

  - Ladder
  - A-J

Digests: 

  - Ladder
  - K091107 (digest 2)
  - K091107 
  - K091146 (digest 2)
  - K091146
  - J23116+J37033 (digest 2)
  - J23116+J37033
  - R0010 (digest 2)
  - R0010
6-30-10.jpg
  • The bands that were cut are marked with a square around them in the picture above. They were also then purified.
  • 3 bands that were cut: 
   - K091107
   - K091146
   - R0010


  • A second gel was ran the same day. The order is provided below: 
  - Ladder
  - B1002
  - Lux R 2
  - C0261+J24679
6-30-10 001.jpg
  • As you can see, the only 2 bands that were cut were B1002 and C0261+J24679


  • Grow for tomorrow: 
  - K274100
  - K112022


July 1

  • Flasks minipreped: 
   - B1006
   - K274100
   - J23100
  • Digests: 
   - B1006 (EX)
   - K274100 (XP)
   - J23100 (SP)
  • Sent to the sequencing center: 
  1) B1002
  2) B1006
  3) S01414
  4) J23100
  5) C0261+J24679


Gel order: 

  - Ladder
  - B1006
  - K274100
  - J23100
7-1-10.jpg
  • All the bands appeared where we wanted them to so we cut out all three bands and then purified them.


  • Ligations: 
  Vector        Inserts

1)J23100 K274100 2)J23100 S01414 3)R0063 a)K274100 4)K091146 b)C0261+J24679 5)K091107 6)R0062 7)R0010 K081007

  • Tomorrow: 
- Develop primers to figure out if K274100 and K112808 are correct.


July 2

  • Colony PCR's ran on gel.The order is as follows: 
   - Ladder
   - 1A, 1B, 1C
   - 2A, 2B, 2C
   - 3A, 3B, 3C

   - Ladder
   - 4A, 4B, 4C
   - 5A, 5B, 5C
   - 6A, 6B, 6C
   - 7A, 7B, 7C
7-2-10.jpg

Unfortunately, we accidentally put ladder in all the Colony products. The picture is provided above.

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