Team:University College London/Week12Joanne2

From 2012.igem.org

(Difference between revisions)
(Created page with "{| class="wikitable" |- ! Gene to be amplified !! Primers |- | irrE || '''Forward primer:''' STF1: 5'-atggggccaaaagctaaagctgaagcc-3' '''Reverse primer:''' ST2R: 5'-tcactgtgcagc...")
 
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ST4R 5'-gtttcttcctgcagcggccgctactagtatcactgtgcagcgtcctgcg-3'
ST4R 5'-gtttcttcctgcagcggccgctactagtatcactgtgcagcgtcctgcg-3'
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|}
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{| class="wikitable"
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|-
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! Component !! 20µl Reaction !! 50µl Reaction !! Final Concentration
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| Nuclease-free water || to 20µl || to 50µl ||
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|-
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| 5X Phusion HF Buffer || 4 µl || 10 µl || 1X
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|-
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| 10 mM dNTPs || 0.4 µl || 1 µl || 200 µM
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|-
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|10 µM Forward Primer || 1 µl || 2.5 µl || 0.5 µM
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|-
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|10 µM Reverse Primer || 1 µl || 2.5 µl || 0.5 µM
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|Template DNA || variable || variable || < 250 ng
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|DMSO (optional) || (0.6 µl) || (1.5 µl) || 3%
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|-
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|Phusion DNA Polymerase || 0.2 µl || 0.5 µl || 1.0 units/50 µl PCR
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Latest revision as of 02:11, 27 September 2012

Gene to be amplified Primers
irrE Forward primer:

STF1: 5'-atggggccaaaagctaaagctgaagcc-3'

Reverse primer:

ST2R: 5'-tcactgtgcagcgtcctgcg-3'

irrE Forward primer:

STF3 5'-gtttcttcgaattcgcggccgcttctagagatggggccaaaagctaaagctgaagcc-3'

Reverse primer:

ST4R 5'-gtttcttcctgcagcggccgctactagtatcactgtgcagcgtcctgcg-3'


Component 20µl Reaction 50µl Reaction Final Concentration
Nuclease-free water to 20µl to 50µl
5X Phusion HF Buffer 4 µl 10 µl 1X
10 mM dNTPs 0.4 µl 1 µl 200 µM
10 µM Forward Primer 1 µl 2.5 µl 0.5 µM
10 µM Reverse Primer 1 µl 2.5 µl 0.5 µM
Template DNA variable variable < 250 ng
DMSO (optional) (0.6 µl) (1.5 µl) 3%
Phusion DNA Polymerase 0.2 µl 0.5 µl 1.0 units/50 µl PCR
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