IGEM:University of East Anglia (UEA), Norwich, UK/2009/Notebook/NRP-UEA-Norwich iGEM/2014/06/30

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Week 3 Day 1

TSL


- Set up 10 Dig-Lig reactions with new BSA1 enzyme provided by TSL.

- DNA was transformed into E.coli using electroporation method previously discussed.

- Transformed bacteria was spreadplated onto canamycin plates containing XGAL and IPTG and incubated O/N at 37°C.

- Mini-prep of DNA obtained from Week 2's successful Dig-Lig experiments according to Qiagen protocol. However, DNA eluted in 25 µl of eluting buffer rather than 50 µl as protocol suggests.

- DNA conc. of each sample quanitifed using the Nanodrop.

- DNA diluted to obtain 5 ng in each 5 µl sample.

- Sense and Antisense primers to each 5 µl DNA (+H20) to achieve 20 µl sample in total and sent for sequencing to verify construct sequence.