IGEM:University of East Anglia (UEA), Norwich, UK/2009/Notebook/NRP-UEA-Norwich iGEM/2013/09/02: Difference between revisions
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A PCR purification was performed - the product was spun down with PB and PE buffers. The nanodrop indicated that tbere was sufficient DNA to work with. <br> | A PCR purification was performed - the product was spun down with PB and PE buffers. The nanodrop indicated that tbere was sufficient DNA to work with. <br> | ||
A restriction digest was performed on pMS82 and the new GUS gene using NdeI and HindIII enzymes in Buffer B. For the GUS gene digest - 5 ul Buffer, 0.5 ul E1 (enzyme 1), 0.5 ul E2, 35 ul template and 9 ul sterile water was used. For the pMS82 digest - 5 ul Buffer, 0.5 ul E1, 0.5 ul E2, 15 ul template and 29 ul sterile water was used. | A restriction digest was performed on pMS82 and the new GUS gene using NdeI and HindIII enzymes in Buffer B. For the GUS gene digest - 5 ul Buffer, 0.5 ul E1 (enzyme 1), 0.5 ul E2, 35 ul template and 9 ul sterile water was used. For the pMS82 digest - 5 ul Buffer, 0.5 ul E1, 0.5 ul E2, 15 ul template and 29 ul sterile water was used. | ||
==Floor Two== | |||
The plates that had been transformed with the ligation (J04450+neo) contained no colonies. | |||
Revision as of 02:13, 4 September 2013
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Floor OneThe PCR of the new GUS gene was performed (8 x 20 ul reactions of same proportions as first time round). Floor TwoThe plates that had been transformed with the ligation (J04450+neo) contained no colonies.
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