Edinburgh/26 June 2008

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==== Thursday 26 June 08 ====
==== Thursday 26 June 08 ====
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* Purified DNA from PCR reactions. Used 20 microlitres of glass beads and eluted to 40 microlitres of TE.<br />
+
* Purified DNA from PCR reactions. Used 20 microlitres of glass beads and eluted to 40 microlitres of TE.
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'''Sample P1:''' ''dxs'' PCR product<br />
+
** Sample P1: ''dxs'' PCR product
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'''Sample P2:''' ''appY'' PCR product<br />
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** Sample P2: ''appY'' PCR product
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'''Sample P3:''' ''glgC'' PCR product<br />
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** Sample P3: ''glgC'' PCR product
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* Set up digests to clone ''appY'' and ''dxs'' into Edinbrick1. Digests with 32μl water, 5μl buffer E, 4μl Edinbrick1 DNA, 4μl purified PCR product, 2.5μl SpeI, 2.5μl EcoRI. Incubated at 37°C. Purified. Set up ligations:<br />
+
* Set up digests to clone ''appY'' and ''dxs'' into Edinbrick1. Digests with 32μl water, 5μl buffer E, 4μl Edinbrick1 DNA, 4μl purified PCR product, 2.5μl SpeI, 2.5μl EcoRI. Incubated at 37°C. Purified. Set up ligations:
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'''Ligation L1:''' ''dsx'' + Edinbrick1, EcoRI/SpeI<br />
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** Ligation '''L1''': ''dsx'' + Edinbrick1, EcoRI/SpeI
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'''Ligation L2:''' ''appY'' + Edinbrick1, EcoRI/SpeI<br />
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** Ligation '''L2''': ''appY'' + Edinbrick1, EcoRI/SpeI
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'''Ligation L3:''' ''glgC'' (5μl) \+linear Babel1 (16-2-8, 2μl) with PNK<br />
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** Ligation '''L3''': ''glgC'' (5μl) + linear Babel1 (16-2-8, 2μl) with PNK
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'''Ligation L4:''' ''glgC'' (5μl) \+linear Babel2 (18-2-8, 2μl) with PNK<br />
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** Ligation '''L4''': ''glgC'' (5μl) + linear Babel2 (18-2-8, 2μl) with PNK<br />
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Incubate ligations at 16 C overnight.
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ligations incubated at 16°C overnight.

Revision as of 14:06, 20 August 2008

Edinburgh iGEM 2008

 

Week 2

Thursday 26 June 08

  • Purified DNA from PCR reactions. Used 20 microlitres of glass beads and eluted to 40 microlitres of TE.
    • Sample P1: dxs PCR product
    • Sample P2: appY PCR product
    • Sample P3: glgC PCR product
  • Set up digests to clone appY and dxs into Edinbrick1. Digests with 32μl water, 5μl buffer E, 4μl Edinbrick1 DNA, 4μl purified PCR product, 2.5μl SpeI, 2.5μl EcoRI. Incubated at 37°C. Purified. Set up ligations:
    • Ligation L1: dsx + Edinbrick1, EcoRI/SpeI
    • Ligation L2: appY + Edinbrick1, EcoRI/SpeI
    • Ligation L3: glgC (5μl) + linear Babel1 (16-2-8, 2μl) with PNK
    • Ligation L4: glgC (5μl) + linear Babel2 (18-2-8, 2μl) with PNK

ligations incubated at 16°C overnight.

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