Difference between revisions of "Team:Munich/recbiobricks.html"
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<h4>Assembling biobrick for RecB, C, D</h4> | <h4>Assembling biobrick for RecB, C, D</h4> | ||
| − | <em>2018/08/31</em> | + | <em>2018/08/31 – 2018/09/07</em> |
<table class="table table-borderless"> | <table class="table table-borderless"> | ||
<tr> | <tr> | ||
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<td>Notes:</td> | <td>Notes:</td> | ||
| − | <td>RecD_BB_EcoRI_fw & RecD_BB_SpeI_rv (3,5 kb) | + | <td> Three PCRs for three BioBricks with primers: RecD_BB_EcoRI_fw & RecD_BB_SpeI_rv (expected: 3,5 kb) |
| − | RecB_BB_EcoRI_fw & | + | RecB_BB_EcoRI_fw & RecB_BB_SpeI_rv (expected: 1,8 kb) |
| − | RecC_BB_EcoRI_fw & | + | RecC_BB_EcoRI_fw & RecC_BB_SpeI_rv (expected: 3,3 kb) |
| − | AT 68° ET 70 s | + | AT 68° ET 70 s; restriction digest of pSB1C3_mRFP with EcoRI and SpeI and PCR purification of the Backbone each PCR was digested with EcoRi and SpeI and ligated into the Backbone. |
| + | |||
</td> | </td> | ||
</tr> | </tr> | ||
Revision as of 16:41, 17 October 2018
Assembling biobrick for RecB, C, D
2018/08/31 – 2018/09/07| Participants: | Enikö Baligács |
| Protocol: | PCR, Agarose gel, Gel extraction, Ligation, Chemical transformation, Mini Prep |
| Notes: | Three PCRs for three BioBricks with primers: RecD_BB_EcoRI_fw & RecD_BB_SpeI_rv (expected: 3,5 kb) RecB_BB_EcoRI_fw & RecB_BB_SpeI_rv (expected: 1,8 kb) RecC_BB_EcoRI_fw & RecC_BB_SpeI_rv (expected: 3,3 kb) AT 68° ET 70 s; restriction digest of pSB1C3_mRFP with EcoRI and SpeI and PCR purification of the Backbone each PCR was digested with EcoRi and SpeI and ligated into the Backbone. |
| Results: | PIC only one worked, but somehow all were ligated :D stuff missing? |
TestPCR for Biobricks of RecB, C, D
2018/09/03| Participants: | Enikö Baligács |
| Protocol: | PCR, Agarose gel |
| Notes: | Primers: VF2 & VR |
| Results: | No results |
Removing PstI restriction site from pSB1C3_RecB
2018/09/03| Participants: | Enikö Baligács |
| Protocol: | PCR, PCR purification, Ligation, Gibson assembly, Transformation |
| Notes: | because of time restraints, this experiment was done using ligation and gibson assembly approaches in parallel PCR: RecB_BB_PstI-remove-Lig Primer: RecB_mut_Lig_fw & RecB_mut_Lig_rv Templ: RecB_BioBrick Plasmid AT: 72° ET: 11 min PCR: RecBCD_BB_PstI_remove_lig Primer: RecB_mut_Lig_fw & RecB_mut_Lig_rv Templ: RecBCD_BioBrick Plasmid AT: 72° ET: 11 min PCR: RecBCD-TEV_BB_PstI_remove-Lig Primer: RecB_mut_Lig_fw & RecB_mut_Lig_rv Templ: RecBCD-TEV_BioBrick Plasmid AT: 72° ET: 11 min PCR: RecB_BB_PstI-remove-GA Primer: RecB_mut_fw & RecB_mut_rv Templ: RecB_BioBrick Plasmid AT: 72° ET: 11 min PCR: RecBCD_BB_PstI_remove_GA Primer: RecB_mut_fw & RecB_mut_rv Templ: RecBCD_BioBrick Plasmid AT: 72° ET: 11 min PCR: RecBCD-TEV_BB_PstI_remove-GA Primer: RecB_mut_fw & RecB_mut_rv Templ: RecBCD-TEV_BioBrick Plasmid AT: 72° ET: 11 min |
| Results: | No results |