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Edinburgh/26 June 2008
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Revision as of 11:04, 20 August 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
Incubate ligations at 16 C overnight.
