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Team:Exeter/lab book/3gip/wk2
From 2012.igem.org
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The 3-Gene Inducible Plasmid: 16th - 20th July 2012 Monday 16th JulyAfternoon • Transformation: RBS Biobrick BBa_B0034, 2012 Distribution Kit Plate 1 2M Tetr_rbs Biobrick BBa_J13002, 2012 Distribution Kit Plate 1 13B Using Invitrogen TOP10 competent cells split into 2 eppendorfs containing 25μl each. Spread on two Ampicillin plates for each transformation using 20μl and 100μls respectively.Tuesday 17th July Afternoon •Adding cultures Cells containing BBa_B0034 and BBa_J13002 were added into liquid medium and incubated overnight Protocol was followed using ampicillin for all BioBrick parts as the selection antibiotic.Wednesday 18th July Morning The incubator had stopped over night and so growth overnight was not optimised. During the miniprep this morning, the cultures were centrifuged with pipette tips in the tubes. The pellets were therefore very small and so the process was repeated overnight using fresh cultures.Afternoon • Adding cultures Cells containing BBa_B0034 and BBa_J13002 were added into liquid medium and incubated overnight Protocol was followed using ampicillin for all BioBrick parts as the selection antibiotic.Thursday 19th July Morning • Mini-Prepping of BBa_B0034 and BBa_J13002 Both BBa_B0034 and BBa_J13002 were nanodropped and recorded low concentrations. This was put down to inexperience with minipreps.• Gel Electrophoresis on a 2% gel was run to check fragment sizes of BBa_B0034 and BBa_J13002 using the EcoR1 and Pst1 enzymes BBa_B0034 showed no band but is only 13bp large and BBa_J13002 showed a very faint band. |
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