Team:Cambridge/Lab book/Week 7

From 2012.igem.org

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'''[[Team:Cambridge/Protocols/PCRProtocol|PCR of Magnesium riboswitch vector fragment B and Magnesium promoter]]'''
'''[[Team:Cambridge/Protocols/PCRProtocol|PCR of Magnesium riboswitch vector fragment B and Magnesium promoter]]'''
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[[File:MgRS vector frag. B.jpg|right|250px|thumb|Riboswitch and promoter gel. Lanes 2+3: Fragment B without 8 codon substitution, lanes 4+5: Fragment B with 8 codon substitution, lanes 6+7: magnesium sensitive promoter, lane 8: +ve control]]
[[File:MgRS vector frag. B.jpg|right|250px|thumb|Riboswitch and promoter gel. Lanes 2+3: Fragment B without 8 codon substitution, lanes 4+5: Fragment B with 8 codon substitution, lanes 6+7: magnesium sensitive promoter, lane 8: +ve control]]
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*Lane 5 accidentally loaded with a DNA ladder instead of loading dye.
*Lane 5 accidentally loaded with a DNA ladder instead of loading dye.
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*Expected fragment sizes:
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:*Lane 2-5: 3kbp
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:*Lane 6-7: 300bp
*After electrophoresis, found vector products had, for the most part, worked. Promoter elements were not amplified - no band of the expected size was observed.
*After electrophoresis, found vector products had, for the most part, worked. Promoter elements were not amplified - no band of the expected size was observed.

Revision as of 10:44, 7 August 2012

Week: 3 4 5 6 7

Contents

Monday

PCR of Magnesium riboswitch vector fragment B and Magnesium promoter


Riboswitch and promoter gel. Lanes 2+3: Fragment B without 8 codon substitution, lanes 4+5: Fragment B with 8 codon substitution, lanes 6+7: magnesium sensitive promoter, lane 8: +ve control
  • Normal PCR settings used, annealing temperature 57 °C, elongation step 90s long.
  • Lane 5 accidentally loaded with a DNA ladder instead of loading dye.
  • Expected fragment sizes:
  • Lane 2-5: 3kbp
  • Lane 6-7: 300bp
  • After electrophoresis, found vector products had, for the most part, worked. Promoter elements were not amplified - no band of the expected size was observed.

Tuesday

Wednesday

Thursday

Friday