Difference between revisions of "Team:Paris Saclay/Notebook/October/10"

(Lab work)
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= Monday 10<sup>th</sup> October=
 
= Monday 10<sup>th</sup> October=
==Lab work==
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===BioBrick K2039000 and K2039001 characterization===
 
===BioBrick K2039000 and K2039001 characterization===
  

Revision as of 15:56, 14 October 2016

Monday 10th October

BioBrick K2039000 and K2039001 characterization

Protein extraction of FRB - GFP 11 in pSB1C3 (pPS16_019) clone 4, FKBP - GFP 10 in pSB1C3 (pPS16_018) clone 6, GFP 1.9 in pUC19 (pPS16_009) clone 1 and PhB1040 strain 594

By Maxence

In order to perform a Western Blot, the proteins were extracted from clones (pPS16_018 clone 6, pPS16_019 clone 4, pPS16_009 clone 1 and PhB1040 strain 594) put on liquide culture the 7th October. For that purpose, the protein extraction protocol was used:

Sample OD (1 ml) Volume of lysis buffer
FRB - GFP 11 in pSB1C3 (pPS16_019) 5.1 127.5 μl
FKBP - GFP 10 in pSB1C3 (pPS16_018) 5.2 130 μl
GFP 1.9 in pUC19 (pPS16_009) 5.3 132.5 μl
PhB1040 strain 594 2.8 70 μl

Protein electrophoresis of FRB - GFP 11 in pSB1C3 (pPS16_019) clone 4, FKBP - GFP 10 in pSB1C3 (pPS16_018) clone 6, GFP 1.9 in pUC19 (pPS16_009) clone 1 and PhB1040 strain 594

By Maxence

A protein electrophoresis of the extracted protein was done using the protein electrophoresis protocol. For that purpose, the gel cassette Bolt 4-12% Bis-Tris Plus 10W was used and the power was set for 1 hour at 65V.

Transfert of protein extracted from FRB - GFP 11 in pSB1C3 (pPS16_019) clone 4, FKBP - GFP 10 in pSB1C3 (pPS16_018) clone 6, GFP 1.9 in pUC19 (pPS16_009) clone 1 and PhB1040 strain 594

By Maxence

The proteins on the electrophoresis gel were transfered on blotting membrane using the protein transfert protocol.