Team:DTU-Denmark/Notebook/29 August 2013

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==Procedure==
==Procedure==
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===Introduce endings for USER cloning in pSB1C3
The table shows the composition of each reaction:
The table shows the composition of each reaction:
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Revision as of 13:34, 29 August 2013

29 August 2013

Contents

lab 208


Main purpose


Who was in the lab


Procedure


===Introduce endings for USER cloning in pSB1C3 The table shows the composition of each reaction:

component 1 2 3 Neg
dNTPs 1uL 1uL 1uL 1uL
HF buffer 10uL 10uL 10uL 10uL
X7 polymerase 0.5uL 0.5uL 0.5uL 0.5uL
MilliQ water 28.5uL 28.5uL 28.5uL 29.5uL
template pSB1C3 1uL 1uL 1uL -
FW primer 57a2 3uL 3uL 3uL 3uL
RV primer 57b 3uL 3uL 3uL 3uL
DMSO (100%) 2uL (4%) 2uL (4%) 2uL (4%) 2uL (4%)
MgCl2 (50mM) 1uL (1mM) 1uL (1mM) 1uL (1mM) 1uL (1mM)

Results


Gel pictures

Ran gels on yesterday's PCRs to create psB1C3 with USER endings.

Small gel:

  • 1 kb ladder
  • 2%, pur
  • 4%, pur
  • 5%, pur
  • 4%, 1uL, pur
  • 4%, 2uL, pur
  • 1M, pur
  • 2%, HQ
  • 4%, HQ
  • 5%, HQ
  • 4%, 1uL, HQ
  • 4%, 2uL, HQ
  • 1M HQ
  • 1 kb ladder (ladder was almost empty so I put another one in the next lane)
  • 1 kb ladder

big gel (samples from touch down PCR):

  • 1 kb ladder
  • GC, HQ
  • 2% HQ
  • 4% HQ
  • 5% HQ
  • 4%, 1uL, HQ
  • 4%, 2uL, HQ
  • 1M HQ
  • GC pur
  • 2%, pur
  • 4%, pur
  • 5%, pur
  • 4%, 1uL, pur
  • 4%, 2uL, pur
  • 1M pur
  • neg
  • 1 kb ladder

Conclusion


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