Team:DTU-Denmark/Notebook/22 August 2013

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22 August 2013

Contents

lab 208


Main purpose


Who was in the lab


Kristian, Henrike

Procedure


USER ligation and transformation

Redid for HAO and cyc in arabinose inducible pZA21::araBAD and for Nir fragments

PCR for Biobrick parts

Set up a new PCR reaction for Biobrick parts using HF buffer and 5% DMSO but no MgCl2. PCR was run on a touchdown program

primers: 53a, 53b

template: Sec2 miniprep

program:

temperature time cycles
98C 2:00 -
98C 0:10 10
63C 1:00 10
-0.5C per cycle
72C 1:00 10
98C 0:10 25
53C 1:00 25
72C 1:00 25
72C 5:00 -
10C hold -

Gel purification

Gel purified the linearized plasmid pZA21::ara with USER endings

colony pPCR to verify AMO insert

Picked 10 colonies from yesterday's cloning for colony PCR. Diluted cells in 50 uL of MilliQ and took 1 uL as template

primers: 53a, 53b

program:

temperature time cycles
98C 10:00 -
98C 0:10 36
56C 0:30 36
72C 2:00 36
72C 5:00 -
10C hold -

nanodrop measurement of ara spl plasmids

Results


Conclusion


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