IGEM:University of East Anglia (UEA), Norwich, UK/2009/Notebook/NRP-UEA-Norwich iGEM/2014/06/25: Difference between revisions
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- Set up the first 11 Dig-Lig experiments, placed into PCR block on the slow protocol. | - Set up the first 11 Dig-Lig experiments, placed into PCR block on the slow protocol. | ||
- Carbenicillin plates made and spreadplated with 40 μl of XGAL and 100 μl of IPTG. | - Carbenicillin plates made and spreadplated with 40 μl of XGAL and 100 μl of IPTG. Plates left to dry upside down in 37°C incubator. | ||
- Transformation of GG DNA into electrocompetent ''E.coli'' cells, PROTOCOL: | |||
• Cells removed from -80°C freezer and allowed to thaw. | |||
• A 5 µl sample of DNA was added to a 50 µl aliquot of cells. | |||
• Transfered each 55 µl sample into a cuvette. | |||
• Electroporation pulse on setting ECR1. | |||
• A 500 µl of SOC broth was added to the cuvette. | |||
• A 555 µl cuvette sample is transferred to an eppendorf tube and incubated for 40 min at 37°C. | |||
Revision as of 06:48, 22 July 2014
 iGEM Project name 1
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Week 2 Day 3TSL - Midi prep following the Qiagen protocol of the 9 constructs previously synthesised by DNA 2.0 and transformed on Day 2 and DNA conc. of each constrcut determined using the nanodrop. Dig-Lig calculation spread sheet completed using these concentrations. - Set up the first 11 Dig-Lig experiments, placed into PCR block on the slow protocol. - Carbenicillin plates made and spreadplated with 40 μl of XGAL and 100 μl of IPTG. Plates left to dry upside down in 37°C incubator. - Transformation of GG DNA into electrocompetent E.coli cells, PROTOCOL: • Cells removed from -80°C freezer and allowed to thaw. • A 5 µl sample of DNA was added to a 50 µl aliquot of cells. • Transfered each 55 µl sample into a cuvette. • Electroporation pulse on setting ECR1. • A 500 µl of SOC broth was added to the cuvette. • A 555 µl cuvette sample is transferred to an eppendorf tube and incubated for 40 min at 37°C.
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