Team:SDU-Denmark/labnotes7
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== Photosensor == | == Photosensor == | ||
+ | |||
+ | === Colony PCR on B0017 === | ||
+ | Date: 27/8<br> | ||
+ | Done by: LC<br> | ||
+ | Methods: PCR<br> | ||
+ | Protocols: CP1.3[https://2010.igem.org/Team:SDU-Denmark/protocols#CP1.3]<br> | ||
+ | Notes: <br> | ||
+ | Premix:<br> | ||
+ | 12,5 µl 10xTAQ Buffer<br> | ||
+ | 5 µl MgCl2 <br> | ||
+ | 5 µl VF2 <br> | ||
+ | 5 µl VR <br> | ||
+ | 2,5 µl dNTP<br> | ||
+ | 17,5 µl H2O<br> | ||
+ | 5/8 µl TAQ Polymerase<br> | ||
+ | <br> | ||
+ | 9,5 µl Premix were added to 15 µl of H2O containing the lysed cells.<br> | ||
+ | <br> | ||
+ | PCR Program:<br> | ||
+ | <table style="text-align: left;" border="1" cellpadding="2" | ||
+ | cellspacing="2"> | ||
+ | <tbody> | ||
+ | <tr> | ||
+ | <td style="vertical-align: top;">Start<br> | ||
+ | </td> | ||
+ | <td style="vertical-align: top;">94 C<br> | ||
+ | </td> | ||
+ | <td style="vertical-align: top;">2 min<br> | ||
+ | </td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td style="vertical-align: top;">Denaturing<br> | ||
+ | </td> | ||
+ | <td style="vertical-align: top;">94 C<br> | ||
+ | </td> | ||
+ | <td style="vertical-align: top;">1 min<br> | ||
+ | </td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td style="vertical-align: top;">Annealing<br> | ||
+ | </td> | ||
+ | <td style="vertical-align: top;">55 C<br> | ||
+ | </td> | ||
+ | <td style="vertical-align: top;">1 min<br> | ||
+ | </td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td style="vertical-align: top;">Elongation<br> | ||
+ | </td> | ||
+ | <td style="vertical-align: top;">72 C<br> | ||
+ | </td> | ||
+ | <td style="vertical-align: top;">30 sec<br> | ||
+ | </td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td style="vertical-align: top;">Goto2<br> | ||
+ | </td> | ||
+ | <td style="vertical-align: top;">rep<br> | ||
+ | </td> | ||
+ | <td style="vertical-align: top;">29x<br> | ||
+ | </td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td style="vertical-align: top;">End<br> | ||
+ | </td> | ||
+ | <td style="vertical-align: top;">72 C<br> | ||
+ | </td> | ||
+ | <td style="vertical-align: top;">3 min<br> | ||
+ | </td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td style="vertical-align: top;">Hold<br> | ||
+ | </td> | ||
+ | <td style="vertical-align: top;">4 C<br> | ||
+ | </td> | ||
+ | <td style="vertical-align: top;"><br> | ||
+ | </td> | ||
+ | </tr> | ||
+ | </tbody> | ||
+ | </table> | ||
+ | <br> | ||
+ | <br> | ||
+ | Results: No useable results, only unclear bands around 120 BP, which seem to be the result of mispriming with VR on B0010. | ||
+ | |||
+ | == Retinal == | ||
==== Futher PCR on POT2 with NinaB (New Primers NO. 5) ==== | ==== Futher PCR on POT2 with NinaB (New Primers NO. 5) ==== |
Revision as of 13:13, 26 September 2010