Team:DTU-Denmark/Notebook/20 August 2013

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===PCR to linearize pZA21::RFP::araBAD===
===PCR to linearize pZA21::RFP::araBAD===
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 +
*Template: pZA21::RFP::araBAD
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*Primers: 55a and 55b
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*Program (A2 in the main machine)- Annealing temperature: 65C
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 +
The table shows the composition of each reaction:
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{| class="wikitable" style="text-align: right"
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! component (per reaction) !! 1 !! 2 !! 3 !! 4 !! 5 !! 6 !! Neg
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|-
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| dNTPs || 1uL || 1uL || 1uL || 1uL || 1uL || 1uL || -
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|-
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| buffer (GC/HF + volume) || HF + 10uL || HF + 10uL || HF + 10uL || GC + 10uL || HF + 10uL || HF + 10uL || HF + 10uL
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|-
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| X7 polymerase || 0.5uL || 0.5uL || 0.5uL || 0.5uL || 0.5uL || 0.5uL || 0.5uL
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|-
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| MilliQ water || 31.5uL || 29uL || 29.5uL || 27uL || 32.5uL
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|-
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| template || 1uL || 1uL || 1uL || 1uL || 1uL || 1uL || -
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|-
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| FW primer 55a || 3uL || 3uL || 3uL || 3uL || 3uL || 3uL || 3uL
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|-
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| RV primer 55b || 3uL || 3uL || 3uL || 3uL || 3uL || 3uL || 3uL
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|-
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| DMSO 100% || 2.5uL (5%) || - || - || 2.5uL (5%) || - || - || -
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|-
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| Betaine 5M || - || 10uL (1M) || - || - || - || - || -
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|-
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|}
==Results==
==Results==

Revision as of 12:32, 20 August 2013

20 August 2013

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Contents

lab 208

Main purpose

  • PCR to linearize pZA21::RFP::araBAD

Who was in the lab

Kristian, Helen, Henrike, Julia

Procedure

PCR to linearize pZA21::RFP::araBAD

  • Template: pZA21::RFP::araBAD
  • Primers: 55a and 55b
  • Program (A2 in the main machine)- Annealing temperature: 65C

The table shows the composition of each reaction:

component (per reaction) 1 2 3 4 5 6 Neg
dNTPs 1uL 1uL 1uL 1uL 1uL 1uL -
buffer (GC/HF + volume) HF + 10uL HF + 10uL HF + 10uL GC + 10uL HF + 10uL HF + 10uL HF + 10uL
X7 polymerase 0.5uL 0.5uL 0.5uL 0.5uL 0.5uL 0.5uL 0.5uL
MilliQ water 31.5uL 29uL 29.5uL 27uL 32.5uL
template 1uL 1uL 1uL 1uL 1uL 1uL -
FW primer 55a 3uL 3uL 3uL 3uL 3uL 3uL 3uL
RV primer 55b 3uL 3uL 3uL 3uL 3uL 3uL 3uL
DMSO 100% 2.5uL (5%) - - 2.5uL (5%) - - -
Betaine 5M - 10uL (1M) - - - - -

Results

Gels

loaded more samples from the screening PCR for BioBrick creation yesterday

  • 1 kb ladder
  • screening D12
  • screening D11
  • screening D10
  • screening D9
  • screening D8
  • screening D7
  • screening D6
  • screening D5
  • screening D4
  • screening D3
  • screening D2
  • screening D1
  • screening C12
  • screening C11
  • screening C10
  • screening C9
  • screening C8
  • 1 kb ladder

(pic big gel)

  • 1 kb ladder
  • screening C7
  • screening C6
  • screening C5
  • screening C4
  • screening C3
  • screening C2
  • screening C1
  • screening B12
  • screening B11
  • screening B10
  • screening B9
  • screening B8
  • screening B7
  • 1 kb ladder

(pic small gel)

Conclusion

Navigate to the Previous or the Next Entry

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