Team:Freiburg/Content/Notebook

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:

Fig.1: Project overview.


Fig.1: Project overview.

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

PCR

Testdigest

p14rz_003

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

PCR

Digest

Testdigest

p14rz_004

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

PCR

PCR

Testdigest

p14rz_005

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

PCR

Digest

Testdigest

p14rz_006

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

PCR

Digest

Testdigest

p14rz_007

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

PCR

Digest

Testdigest

p14rz_008

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

PCR

Testdigest

p14rz_009

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

Digest

Testdigest

p14rz_010

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

PCR

PCR

Testdigest

p14ls_003

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

PCR

Digest

Testdigest

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

Digest

Digest

Testdigest

p14vv_002

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

PCR

Digest

Testdigest

p14vv_003

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

PCR

Testdigest

p14vv_006

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

PCR

Testdigest

p14vv_007

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

PCR

Testdigest

p14vv_008

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

PCR

Digest

Testdigest

p14vv_009

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

PCR

Digest

Testdigest

p14vv_010

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

PCR

Digest

Testdigest

p14vv_012

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

PCR

Digest

Testdigest

p14vv_013

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

PCR

Digest

Testdigest

p14vv_014

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

PCR

Digest

Testdigest

p14zl_010

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

PCR

Digest

Testdigest

p14zl_011

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

PCR

Digest

Testdigest

p14zl_016

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

PCR

Digest

Testdigest

p14zl_017

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

PCR_digest

Testdigest

p14zl_020

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

PCR

Digest

Testdigest