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From 2011.igem.org

August 28th - September 3rd

Sunday, August 28

Monday, August 29

Tuesday, August 30

Afternoon - Evening lab work done by: Claire Paduano, Youjin Cho, Maneesh Gupta, James Mathew, Charlie Chung

• Digestion Reactions

- specifics to be posted soon

• Agarose Gel Electrophoresis

- specifics to be posted soon

• Gel Extraction and Purification of DNA

- specifics to be posted soon (including some issues in cutting out the insert band vs. backbone band)

• NanoDrop Spectrophometry for Purified DNA Quantification

- specific [ ]s to be posted soon

• Ligation Reactions

- specifics to be posted soon

Wednesday, August 31

Afternoon/Evening Lab work done by: Claire Paduano and Maneesh Gupta

• Looked at ATTO590 coated chip from Aug 27th under fluorescent microscope: channels still fluorescent

Photo to be posted soon

Streptavidin coating and Fluorescent Probes

Procedure to be posted soon

Flow experiments

Details and photos to be posted soon

Thursday, September 1

Morning lab work done by: Claire Paduano and Nancy Li

• Coated three chips with streptavidin

Streptavidin coating

• Protocol from Gleghorn et al: http://www.ncbi.nlm.nih.gov/pubmed/20024046

1) 45 min in 4% (by volume) MPTMS in ethanol

2) 20 min in 1mM GMBS

3) 45 min in 25ng/mL NeutrAvidin in PBS

Evening lab work done by: Maneesh Gupta

• Did continuous flow experiments to test resilience of biotin-avitin binding
• Used same exposure (83.33) in all images taken

Friday, September 2

Afternoon lab work done by: Charlie Chung

• Miniprep DNA purification of cultures containing pSB1C3 vector backbone with (RFP+Avi-Tag), (vioA+Avi-Tag), (vioB+Avi-Tag), and (vioE+Avi-Tag) inserts -- in preparation for iGEM BioBrick submission

Saturday, September 3

Afternoon Lab accomplished by: Jim Mathew, Charlie Chung, Nancy Li

Miniprep -took concentrations of yesterday miniprep -all 260/280nm ratio were precisely near 1.8 indicting pure DNA

DNA Concentration (ng/uL)

RFP 1 in PSB1C3 Backbone 344.0 RFP 2 in PSB1C3 Backbone 256.4 RFP 3 in PSB1C3 Backbone 230.4 VIOA 1-1 in PSB1C3 Backbone 205.9 VIOA 1-2 in PSB1C3 Backbone 494.3 VIOA 1-3 in PSB1C3 Backbone 370.8 VIOA 2-1 in PSB1C3 Backbone 892.5 VIOA 2-2 in PSB1C3 Backbone 203.8 VIOA 2-3 in PSB1C3 Backbone 206.0 VIOB 1-1 in PSB1C3 Backbone 503.9 VIOB 1-2 in PSB1C3 Backbone 186.1 VIOB 1-3 in PSB1C3 Backbone 398.8 VIOB 2-1 in PSB1C3 Backbone 868.9 VIOB 2-2 in PSB1C3 Backbone 145.6 VIOB 2-3 in PSB1C3 Backbone 174.6 VIOE 1 in PSB1C3 Backbone 266.2 VIOE 2 in PSB1C3 Backbone 552.9 VIOE 3 in PSB1C3 Backbone 400.7

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