IGEM:University of East Anglia (UEA), Norwich, UK/2009/Notebook/NRP-UEA-Norwich iGEM/2013/09/02: Difference between revisions
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|style="background-color: #EEE"|[[Image:igem-logo-150px.png|150px]]<span style="font-size:22px;"> iGEM Project name 1</span> | |style="background-color: #EEE"|[[Image:igem-logo-150px.png|150px]]<span style="font-size:22px;"> iGEM Project name 1</span> | ||
|style="background-color: #F2F2F2" align="center"| | |style="background-color: #F2F2F2" align="center"|[[File:Report.png|frameless|link={{#sub:{{FULLPAGENAME}}|0|-11}}]][[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|[[File:Resultset_previous.png|frameless|link={{#lnpreventry:{{FULLPAGENAME}}}}]][[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]] }}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]][[File:Resultset_next.png|frameless|link={{#lnnextentry:{{FULLPAGENAME}}}}]]}} | ||
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==Floor One== | ==Floor One== | ||
[[Image:GEL3.png|thumb| Fig 1. Gel showing another GUS PCR, with a 1Kb+ base ladder in lane 2 and a band of GUS product at roughly 1.8Kb in lane 3.]] | |||
The PCR of the new GUS gene was performed (8 x 20 ul reactions of same proportions as first time round).<br> | 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> | A 1% agarose gel was made.<br> | ||
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Performed some streak purifications and lawns.<br> | Performed some streak purifications and lawns.<br> | ||
Made 2 x 500 ml SFM (+ Ny).<br> | Made 2 x 500 ml SFM (+ Ny).<br> | ||
An agarose | An agarose gel electrophoresis of the PCR products showed a fragment at approx. 1.8 Kb which is likely to be the GUS gene ''fig 1''.<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 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. | |||
Latest revision as of 23:15, 26 September 2017
iGEM Project name 1 | Main project page Previous entry Next entry |
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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