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From 2014.igem.org

Cloning

The protocols for our methods can be found here.

During our experiments, we conducted a lot of PCRs with different Primers. In order to stay on top of things, we numbered Primers, PCRs and constructs. You can download both lists with detailed information about Primer composition and about PCRs in the following documents:

p14rz_002

• PCR: o14rz_005, o14_rz006; Template pQXCIN-SLC7A1
• Digest: pKM006, EcoRI + NotI; PCR-product
• Testdigest with PstI in Cutsmart, expected bands: 5,9; 0,7

p14rz_003

• PCR 3
• Digest 3, 4
• Testdigest with PstI + SpeI in Cutsmart, expected bands: 4,1;2,1;1,8

p14rz_004

• PCR 21,22
• Digest -
• Testdigest with PstI in Cutsmart, expected bands: 4,1;3,2

p14rz_005

• PCR 4,5
• Digest 5
• Testdigest with AgeI+HindIII in Cutsmart, expected bands: 6,6;1,3

p14rz_006

• PCR 17,18
• Digest 5
• Testdigest with AgeI+HindIII in Cutsmart, expected bands: 6,6;1,4

p14rz_007

• PCR 37,38
• Digest 21
• Testdigest with PstI in Cutsmart, expected bands: 4,1;2,2;1,7

p14rz_008

• PCR blue light receptor; Template: pQXCIN-SLC7A1
• Digest pKM006 EcoRI, NotI
• Testdigest with SmaI in Cutsmart, expected bands: 5,7

p14rz_009

• PCR 39,40
• Digest 5
• Testdigest with BamHI in Cutsmart, expected bands: 3,0;2,7

p14rz_010

• PCR 23,24
• Digest 12
• Testdigest with PstI in Cutsmart, expected bands:3,1;2,3;0,56

p14ls_003

• PCR 7,9,10
• Digest -
• Testdigest with KpnI in Cutsmart, expected bands: 4,9; 1,3; 0,4

p14vv_001

• PCR Primers vv003,vv004; Template: pMIG
• Digest pMIG EcoRI,NotI; C1-eGFP EcoRI,NotI
• Testdigest with NgoMIV in Cutsmart, expected bands: 5,7; 0,6

p14vv_002

• PCR -
• Digest 8,11
• Testdigest with PstI in Cutsmart, expected bands: 3,2;2,1;1,4

p14vv_003

• PCR pMIG with primer vv003+vv004-
• Digest xx
• Testdigest with KpnI in Cutsmart, expected bands: 3,5;1,5

p14vv_006

• PCR 26
• Digest 13
• Testdigest with NgoMIV in Cutsmart, expected bands: 3,5;1,9;0,95,0,6

p14vv_007

• PCR 27
• Digest 14
• Testdigest with NgoMIV in Cutsmart, expected bands: 3,5; 1,3; 0,97; 0,66

p14vv_008

• PCR 43
• Digest 22
• Testdigest with EcoRV in Cutsmart, expected bands: 3,4; 1,4 0,75

p14vv_009

• PCR 44
• Digest 22
• Testdigest with EcoRV+MluI in Cutsmart, expected bands: 3,4; 1,43; 0,75

p14vv_010

• PCR 45
• Digest 22
• Testdigest with EcoRV in Cutsmart, expected bands: 3,4; 2,2

p14vv_012

• PCR 46
• Digest 22
• Testdigest with EcoRV in Cutsmart, expected bands: 4,7;3,4

p14vv_013

• PCR 47,48,49
• Digest 22
• Testdigest with NgoMIV in Cutsmart, expected bands: 3,4;2,0;1,3;0,2

p14vv_014

• PCR 50,51
• Digest 22
• Testdigest with NgoMIV in Cutsmart, expected bands: 4,1; 1,56; 0,66

p14zl_010

• PCR 1,2
• Digest 1
• Testdigest with NgOMIV in Cutsmart, expected bands: 4,7;2,4;1,1

p14zl_011

• PCR 19,20
• Digest 1
• Testdigest with NgOMIV in Cutsmart, expected bands: 4,7;2,0;1,3;0,43

p14zl_016

• PCR 52,53
• Digest 22
• Testdigest with HindIII in Cutsmart, expected bands: 5,7;2,3;1,7

p14zl_017

• PCR 64,65
• Digest 24
• testdigest with NgoMIV in Cutsmart, expected bands: 5,4;2,9

p14zl_020

• PCR 59,60
• Digest 24
• testdigest with BamHI-HF, NgoMIVin Cutsmart, expected bands: 4,7; 2,4; 1,8