Week of 8/2

==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)



[https://2010.igem.org/Team:Penn_State/Notebook Return to Notebook]