Team:Stockholm/30 August 2010

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(Andreas)
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{{Stockholm/Top2}}
{{Stockholm/Top2}}
==Andreas==
==Andreas==
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===Cloning of SOD into pMA.His===
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====Transformation results====
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''From 28 28/8''
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Good colony yield. Four colonies (SH1-SH4) picked for colony PCR.
 +
 +
====Colony PCR====
 +
*SH1-SH4: pMA.SOD⋅His
 +
*PC: Positive control; pMA.His
 +
*NC: Negative control; blank
 +
Procedures according to standard colony PCR protocol. Elongation time 1:00.
 +
 +
====Gel verification====
 +
[[image:ColPCR_pMA.SOD*His_30aug.png|200px|thumb|right|'''Gel verification of SOD cloning into pMA.His.'''<br />3 &mu;l &lambda;, 4 &mu;l sample.<br />&lambda;=O'GeneRuler 1 kb DNA ladder.]]
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1 % agarose, 100 V
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Expected bands:
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*'''pMA.SOD&sdot;His:''' 831 bp
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*'''pMA.His:''' 348 bp
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 +
''Results''<br />
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Well corresponding bands indicating successful insertion of SOD into the vector.
 +
 +
====ON cultures====
 +
SH1 and SH2 selected for plasmid prep and sequencing. Set 5 ml LB + 100 Amp ON cultures. 37 &deg;C, 225 rpm.

Revision as of 07:49, 31 August 2010


Contents

Andreas

Cloning of SOD into pMA.His

Transformation results

From 28 28/8 Good colony yield. Four colonies (SH1-SH4) picked for colony PCR.

Colony PCR

  • SH1-SH4: pMA.SOD⋅His
  • PC: Positive control; pMA.His
  • NC: Negative control; blank

Procedures according to standard colony PCR protocol. Elongation time 1:00.

Gel verification

Gel verification of SOD cloning into pMA.His.
3 μl λ, 4 μl sample.
λ=O'GeneRuler 1 kb DNA ladder.

1 % agarose, 100 V

Expected bands:

  • pMA.SOD⋅His: 831 bp
  • pMA.His: 348 bp

Results
Well corresponding bands indicating successful insertion of SOD into the vector.

ON cultures

SH1 and SH2 selected for plasmid prep and sequencing. Set 5 ml LB + 100 Amp ON cultures. 37 °C, 225 rpm.