Team:Stockholm/17 August 2010

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==Andreas==
==Andreas==
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===Site-directed mutagenesis===
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''Continued from 16/8''
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 +
====Glycerol stocks====
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''From 16/8 ON cultures''
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 +
yC1: pSB1C3.m.yCCS 1, 17/8
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yC4: pSB1C3.m.yCCS 4, 17/8
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SC1: pSB1C3.mut SOD 1, 17/8
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SC2: pSB1C3.mut SOD 2, 17/8
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 +
1600 μl cell culture in 400 μl 100 % glycerol
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====Plasmid prep====
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{|border="1" cellpadding="2" cellspacing="0" align="right"
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!colspan="5"|DNA concentrations
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|-
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! 
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!colspan="2"|Prior to concentration
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!colspan="2"|After concentration
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|-
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!Sample
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!Conc [ng/μl]
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!A<sub>260</sub>/A<sub>280</sub>
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!Conc [ng/&mu;l]
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!A<sub>260</sub>/A<sub>280</sub>
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|-
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|yC1&rarr;pSB1C3.m-yCCS 1
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|36.88
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|1.82
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|250.2
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|1.92
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|-
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|yC4&rarr;pSB1C3.m-yCCS 4
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|41.83
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|1.77
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|215.7
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|1.87
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|-
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|SC1&rarr;pSB1C3.m-SOD 1
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|47.38
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|1.72
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|230.2
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|1.91
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|-
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|SC2&rarr;pSB1C3.m-SOD 2
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|38.00
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|1.72
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|104.0
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|1.86
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|}
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''From 16/8 ON cultures''
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 +
Elution volume: 70 &mu;l dH<sub>2</sub>O
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 +
Since samples were intended for sequencing, they had to be concentrated in the evaporator. Results in table.<br />
 +
Samples sent for sequencing from VF2 primer annealing site:
 +
*15 &mu;l DNA
 +
*1.5 &mu;l 10 mM VF2 primer
 +
 +
====ON cultures====
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New ON cultures (5 ml + 25 Cm) were set for plasmid prep from same clones as were sent for sequencing: yC1, yC4, SC1, SC2.

Revision as of 15:22, 18 August 2010

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Contents

Andreas

Site-directed mutagenesis

Continued from 16/8

Glycerol stocks

From 16/8 ON cultures

yC1: pSB1C3.m.yCCS 1, 17/8 yC4: pSB1C3.m.yCCS 4, 17/8 SC1: pSB1C3.mut SOD 1, 17/8 SC2: pSB1C3.mut SOD 2, 17/8

1600 μl cell culture in 400 μl 100 % glycerol

Plasmid prep

DNA concentrations
  Prior to concentration After concentration
Sample Conc [ng/μl] A260/A280 Conc [ng/μl] A260/A280
yC1→pSB1C3.m-yCCS 1 36.88 1.82 250.2 1.92
yC4→pSB1C3.m-yCCS 4 41.83 1.77 215.7 1.87
SC1→pSB1C3.m-SOD 1 47.38 1.72 230.2 1.91
SC2→pSB1C3.m-SOD 2 38.00 1.72 104.0 1.86

From 16/8 ON cultures

Elution volume: 70 μl dH2O

Since samples were intended for sequencing, they had to be concentrated in the evaporator. Results in table.
Samples sent for sequencing from VF2 primer annealing site:

  • 15 μl DNA
  • 1.5 μl 10 mM VF2 primer

ON cultures

New ON cultures (5 ml + 25 Cm) were set for plasmid prep from same clones as were sent for sequencing: yC1, yC4, SC1, SC2.

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