Team:Stanford/Notebook/Lab Work/Week 5

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Contents

7/19 Monday

Laura's Lab Notebook

set up the following ligations from the gel extractions done by me, Karina, and Francisco on Friday, 7/14/10)

  • Francisco ran and imaged diagnostic gel on Friday
Ligation Recipe
dH2O none
vector (with terminators attached) 5.0 uL
insert (RFP or GFP) 12.0 uL
10X buffer 2.0 uL
T4 ligase 1.0 uL
  • normally run for 10 minutes at room temperature, overnight this time (started at 11:30 am)


Nanodrop data (deemed unreliable based on 260/230, possibly due do residual EtBr contamination)

part 260/280 260/230 ng/uL
vector/terminator 1.77 0.03 19.1
RFP 1.86 0.02 16.2
GFP 1.83 0.02 14.8

7/20 Tuesday

Laura's Lab Notebook

helped Alex with preparation of competent cells

  • see protocol: [http://openwetware.org/wiki/Stanford/BIOE44:Module_1:Day3 Preparing Electrocompetent Cells]


ran diagnostic gel for PCR-assembled DNA (Karina set up PCR rxns yesterday)

  • order on gel:
  1. 100 bp ladder
  2. RSID1/RBS
  3. RSID2/RBS