Team:Munich/recbiobricks.html

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