Team:Exeter/lab book/3gip/wk2

Operon Construction  |  Glycobase

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Afternoon

•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

•Transferring colonies to liquid medium

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 stopped unexpectedly during the night, this meant growth was not optimised. During the miniprep this morning, the cultures were centrifuged with pipette tips in the tubes. The pellets were very small and therefore the process was repeated overnight using fresh cultures.

Afternoon

•Transferring colonies to liquid medium

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.

Morning

•Miniprepping of BBa_B0034 and BBa_J13002

• 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.