Team:Stockholm/12 October 2010

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{{Stockholm/Top2}}
{{Stockholm/Top2}}
==Andreas==
==Andreas==
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===Removal of insertion in BioBrick suffixes===
 +
====Transformation results====
 +
Good colony yield on all plates. A few red colonies discovered on all plates, indicating that some partly digested pSB1C3 vector, still containing the RFP insert, had been excised during gel extraction.
 +
 +
====Colony PCR====
 +
''From 11/10 transformations'''
 +
 +
#pSB1C3.IgGp A-C
 +
#pSB1C3.bFGF A-C
 +
#pSB1C3.ProtA A-C
 +
#pSB1C3.yCCS A-C
 +
#pSB1C3.SOD A-C
 +
 +
*Standard colony PCR settings
 +
**1:15 elongation time
 +
 +
====Gel verification====
 +
[[image:ColPCR_reminserts_gel1_12oct.png|200px|thumb|right|'''Gel 1. Colony PCR gel verification of cloned with insertion removed.'''<br />3 &mu;l &lambda;; 5 &mu;l sample.<br />&lambda; = O'GeneRuler 1 kb DNA ladder.]]
 +
[[image:ColPCR_reminserts_gel2_12oct.png|200px|thumb|right|'''Gel 2. Colony PCR gel verification of cloned with insertion removed.'''<br />3 &mu;l &lambda;; 5 &mu;l sample.<br />&lambda; = O'GeneRuler 50 bp DNA ladder.]]
 +
'''Gel 1:''' 1 % agarose, 140 V<br />
 +
'''Gel 2:''' 1 % agarose, 160 V
 +
 +
'''Expected bands'''
 +
#pSB1C3.IgGp: 1262 bp
 +
#pSB1C3.bFGF: 794 bp
 +
#pSB1C3.ProtA: 506 bp
 +
#pSB1C3.yCCS: 1076 bp
 +
#pSB1C3.SOD: 791 bp
 +
 +
'''Results'''
 +
#All clones correct.
 +
#All clones correct.
 +
#All clones correct.
 +
#Correct size for clone C, weak band for clone B. Clone A probably carrying an RFP insert.
 +
#All clones correct.

Revision as of 14:20, 14 October 2010

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Contents

Andreas

Removal of insertion in BioBrick suffixes

Transformation results

Good colony yield on all plates. A few red colonies discovered on all plates, indicating that some partly digested pSB1C3 vector, still containing the RFP insert, had been excised during gel extraction.

Colony PCR

From 11/10 transformations'

  1. pSB1C3.IgGp A-C
  2. pSB1C3.bFGF A-C
  3. pSB1C3.ProtA A-C
  4. pSB1C3.yCCS A-C
  5. pSB1C3.SOD A-C
  • Standard colony PCR settings
    • 1:15 elongation time

Gel verification

Gel 1. Colony PCR gel verification of cloned with insertion removed.
3 μl λ; 5 μl sample.
λ = O'GeneRuler 1 kb DNA ladder.
Gel 2. Colony PCR gel verification of cloned with insertion removed.
3 μl λ; 5 μl sample.
λ = O'GeneRuler 50 bp DNA ladder.

Gel 1: 1 % agarose, 140 V
Gel 2: 1 % agarose, 160 V

Expected bands

  1. pSB1C3.IgGp: 1262 bp
  2. pSB1C3.bFGF: 794 bp
  3. pSB1C3.ProtA: 506 bp
  4. pSB1C3.yCCS: 1076 bp
  5. pSB1C3.SOD: 791 bp

Results

  1. All clones correct.
  2. All clones correct.
  3. All clones correct.
  4. Correct size for clone C, weak band for clone B. Clone A probably carrying an RFP insert.
  5. All clones correct.
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