Team:Warsaw/Calendar-Main/8 August 2009
From 2009.igem.org
Assembly of endosomal detection operon
Marcin
Task 1:
- Assembly of BBa_B0032 - cro construct
- Gel electrophoresis and subsequent purification of digested products via gel-out
Evaluation of results of the digestion
Procedure:
- Fragments of agarose gel were carefully cut out and in the next step DNA was extracted from the gel using the A&A gel-out kit. Detailed procedure is described here
Task 2:
- Cloning cro coding sequence to pSB1A3 vector
Methods:
- Ligation mixture composition:
16 μl digested cro 10 μl digested vector 3 μl ligation buffer (Fermentas) 1 μl ligase T4 (Fermentas)
- Duration of ligation was about 18 hours; reaction was conducted in 19 °c (approximately).
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