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Team:Toronto/Notebook/31 August 2011
From 2011.igem.org
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J23100+B0034+C0012+J04500 construct samples 22 / 24 / 14 were digested with SpeI and PstI for the insertion of His-5 MMS6.
MMS6 was also PCR amplified for this insertion. A duplicate reaction was performed in case of downstream reactions failed
A gel was run to confirm the digestion and that the PCR reaction produced only one band.
<insert image of gel Aug 31 (1)> --> Suraj has chart of lane contents.
From the gel it was confirmed that the PCR had no side reactions and that the J23100 construct was cut
The PCR products were then purified using the Geneaid Kit
PCR reaction 1 (p) when eluted in 50ul of elution buffer had a concentration of 23.5ng/ul'
PCR reaction 1 (g) when eluted in 50ul of elution buffer had a concentration of 41.4ng/ul'
The J23100 digested constructs were then Gel purified. resulting in a concentration of - add when completed
An overnight ligation was then performed.
