"

From 2010.igem.org

Contents 1 August 2nd 2 August 3rd 3 August 4th 4 August 5th 5 August 6th

August 2nd

• Miniprep

- R0061
- R1062
- J23100 

• Digests

- K091146 (SP)
- R1062 (SP)
- J23100 (SP)
- K091107 (SP)
- Lux+RBS+A3 (XP+phosphatase)
- RFP+RBS (XP+phophatase)

• Gel order:

- J23100
- R1062
- K091146
- R0063
- K091107
- K145150
- Ladder

• Ligations

- J23100 + Lux+RBS+A3
- R1062 + RFP+RBS
- K091146 + RFP+RBS
- K091107 + RFP+RBS

August 3rd

• Colony PCR

1)K091107+RFP+RBS A-C
2)R1062+RFP+RBS A-D
3)J23100+LuxR+RBS+A3 A-D

• Digest

- R0063 (SP)
- K145150 (SP)
- K091146 (SP)
- RFP+RBS (XP+phosphatase)
- Violet pigment 1 (EP)
- Violet pigment 2 (EP)
- A3 (EP+phosphatase)

• Digest for 6 hours:

- J24679+B1002 (B)
- J24679+B1002 (D)

• Gel order:

- Violet 1
- Violet 2
- K145150 
- R0063
- K091146 
- Ladder

The last two should have been about 7000 base pairs but it was much lower so the pieces were not cut out. The first piece (K091146) was cut out.

• Gel number # 2:

-Row 1
  - Ladder
  - J24679+B1002 (B)
  - J24679+B1002 (D)
Row 2
  - Ladder
  - 1A 
  - 1B
  - 1C
  - 2A
  - 2B
  - 2C
  - 2D
  - 3A
  - 3B
  - 3C
  - 3D

• PCRs did not show up. But, the bands for J24679+B1002 (B) and (D) were extracted

August 4th

• Miniprep:

- R0061

• Digests

- R0061 (SP+phosphatase)
- R0062 (SP+phosphatase)
- R0063 (SP+phosphatase)
- K145150 (SP+phosphatase)
- K091146 (SP+phosphatase)
- RFP+RBS (XP)
- J23100 (SP+phosphatase)
- Lux+RBS (XP)

• Grow Up for tomorrow:

- RFP+RBS
- K091107
- K091146

• Sent to sequencing:

- K091146+RFP+RBS (A-D)

August 5th

• Digests

- RFP (E)
- RFP (X)
- RFP (S)
- RFP (P)
- RFP (EP)
- RFP (XS)
- Lux promoters with RFP but used violet pigment as the backbone

• Minipreps:

- K091146+RFP+RBS
- RFP+RBS
- K091107
- K091146

• Ligations:

- R0061 + RFP+RBS
- R0063 + RFP+RBS
- K091146 + RFP+RBS
- K145150 + RFP+RBS
- K091107 + RFP+RBS
- R1062 + RFP+RBS

• Colony PCR

- J23100+Lux+RBS (A-F)

• Gel order:

- Ladder
- J23100+Lux+RBS A-F
- Ladder 2
- RFP (E)
- RFP (X)
- RFP (S)
- RFP (P)
- RFP (EP)
- RFP (XS)

August 6th

• Miniprep

-RFP+RBS

• Gel order of PCR products:

- Ladder        - Ladder 2
- 1A            - 4B
- 1B            - 4C
- 1C            - 4D 
- 1D            - 5A
- 2A            - 5B
- 2B            - 5C
- 2C            - 6A
- 2D            - 6B
- 3A            - 6C
- 3B 
- 3C
- 3D
- 4A

• Colony PCR

- R0063+RFP (A-D)
- K091107+RFP (A-D)
- K145150+RFP (A-D)
- R0061+RFP (A-D)
- K091146+RFP (A-C)
- R1062+RFP (A-C)

• Today, we compared our enzymes to Mike Speer's (grad student) enzymes because we aren't sure if they are working correctly.

• Sent to sequencing:

1)K091107+RFP (a)
2)R0061+RFP (a)

Return to Notebook