Team:SDU-Denmark/labnotes11
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= Lab notes (9/20 - 9/26) = | = Lab notes (9/20 - 9/26) = | ||
__TOC__ | __TOC__ | ||
+ | |||
+ | == Photosensor group == | ||
+ | === Miniprep of PS (composit part) in pSB3T5 === | ||
+ | '''Date:''' 9/22 2010<br> | ||
+ | '''Done By:''' Lc and Maria<br> | ||
+ | '''Protocol:''' [https://2010.igem.org/Team:SDU-Denmark/protocols#MP1.2 MP1.2]<br> | ||
+ | '''Notes:''' <br> | ||
+ | col. N and J from the coloni PCR was selected and miniprep'ed according to protocol. Sample 1 of each colni was eluted in 20uL elution buffer, and sample 2 of each coloni was eluted according to protocol.<br> | ||
+ | To ensure thet the plasimids contained the correct insert, the miniprep'ed plasmids were digested with EcoRI and PstI.<br> | ||
+ | Restriction mixture:<br> | ||
+ | 12uL H2O<br> | ||
+ | 2uL FD green buffer<br> | ||
+ | 1uL EcoRI<br> | ||
+ | 1uL PstI<br> | ||
+ | 5uL Plasmid from either col. N or J<br> | ||
+ | |||
+ | The samples are digested for 10 min. at 37C, and samples are loaded onto a 1% agarose gel. Undigested plasmid is used as controle and gene ruler DNA ladder mix is used as marker.<br> | ||
+ | Loading scheme:<br> | ||
+ | <table style="text-align: left;" border="1" cellpadding="2" cellspacing="2"> | ||
+ | <tr> | ||
+ | <td>Lanes</td> | ||
+ | <td>Sample</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>1</td> | ||
+ | <td>marker</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>2</td> | ||
+ | <td>col. J </td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>3</td> | ||
+ | <td>col J (uncut)</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>4</td> | ||
+ | <td>col N</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>5</td> | ||
+ | <td>col N (uncut)</td> | ||
+ | </tr> | ||
+ | </table><br><br> | ||
+ | '''Results:'''<br><br> | ||
+ | '''Analysis:'''<br> | ||
+ | Two bands are observed en the lanes of the digestes samples representing undigested plasmid and insert.<br> | ||
+ | Miniprep samples are sent for sequencing and is stored at -20C.<br> | ||
+ | --[[User:Tipi|Tipi]] 11:58, 30 September 2010 (UTC)<br><br> | ||
== Flagella Group == | == Flagella Group == | ||
- | === | + | === Restriction Digest of FlhDCmut-B0015 and pSB3T5 with J13002 === |
+ | |||
+ | '''Date:''' 9/25 2010<br> | ||
+ | '''Done By:''' Louise, Pernille and Sheila<br> | ||
+ | '''Protocol:''' [https://2010.igem.org/Team:SDU-Denmark/protocols#RD1.1 RD1.1] and [https://2010.igem.org/Team:SDU-Denmark/protocols#DE1.3 DE1.3]<br> | ||
+ | '''Notes:''' <br> | ||
+ | '''Results:''' gel extraction<br> | ||
+ | '''Conclusion:''' <br><br> | ||
+ | |||
+ | === Ligation of pSB3T5 with J13002 and FlhDCmut-B0015 === | ||
+ | |||
+ | '''Date:''' 9/25 2010<br> | ||
+ | '''Done By:''' Louise, Pernille and Sheila<br> | ||
+ | '''Protocol:''' [https://2010.igem.org/Team:SDU-Denmark/protocols#LG1.2 LG1.2]<br> | ||
+ | '''Notes:''' Samples were prepared in the ratios 1:1, 1:2 and 1:4<br> | ||
+ | '''Results:''' <br> | ||
+ | '''Conclusion:''' <br><br> | ||
+ | |||
+ | === Transformation of FlhDCmut-B0015 inserted into pSB3T5 with J13002 === | ||
'''Date:''' 9/26 2010<br> | '''Date:''' 9/26 2010<br> |
Latest revision as of 11:58, 30 September 2010