IGEM:University of East Anglia (UEA), Norwich, UK/2009/Notebook/NRP-UEA-Norwich iGEM/2013/09/02: Difference between revisions
(Autocreate 2013/09/02 Entry for IGEM:University_of_East_Anglia_(UEA),_Norwich,_UK/2009/Notebook/NRP-UEA-Norwich_iGEM) |
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== | ==Floor One== | ||
The PCR of the new GUS gene was performed (8 x 20 ul reactions of same proportions as first time round).<br> | |||
A 1% agarose gel was made.<br> | |||
Took photographs of all Bioassay plates where actinomycete colonies appeared to have a zone of clearing around them - indicating the production of an anti-fungal.<br> | |||
Spore stocks were filtered, concentrated and resuspended in 20 % glycerol.<br> | |||
Isolated Streptomyces sp. colonies from the latest soil sample plates and streaked onto new plates. <br> | |||
Performed some streak purifications and lawns.<br> | |||
Made 2 x 500 ml SFM (+ Ny).<br> | |||
An agarose fel electrophoresis of the PCR products showed a fragment at approx. 1.8 Kb which is likely to be the GUS gene.<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. | |||
Revision as of 04:49, 3 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).
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