Team:Hawaii/Notebook/2008-10-23

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Things we did today

Wetlab work

Antibiotic test

Grace

No change from yesterday
Colony PCR of 6 colonies from sp_{40, 50, 75} and 8 colonies from sp₁₀₀ plates

sp_75,100 colonies correct

Restreaked sp_2.5 blue colony, sp₁₀₀ #1 & 2 on LB+amp₁₀₀, LB, LB+sp₁₀₀ + X-gal

Large scale plasmid prep

Grace

From sp₁₀₀ plate: #1, 2
From 9/28 sp_2.5 plate: #1

Gel showed no DNA in lanes migrated. Huh? Will try one more time to plasmid prep tomorrow.

Inoculated for plasmid prep

Grace

In 10 ml TB+amp₁₀₀:

nir
nir+rbs
slr1
pilA
GFPf
slr1+GFPf

In 50 ml TB+sp₁₀₀:

BB1 (sp₁₀₀ plate)

Incubated at 37C for 18 hours with shaking at 250 rpm

Verification of transformants

EtBr stained 2% agarose gel ran at 95V for 1 hour. Two microliters of PCR product were loaded into each lane.

Grace

**Colony count of transformants**
Plasmid	Colonies
J04430	0
(-) on amp₁₀₀	0
BB-1 (plasmid from 9/28 plate)	4
(-) on sp₁₀₀	0

Colony PCR of 4 BB-1 transformants

None were correct

RE digest

Grace

EcoRI/PstI: nir+rbs
XbaI/SpeI: slr1+GFPf

Plasmid Prep

Krystle

Plasmid prep of
- lac+rbs+slr+gfpf #1,3,4
- nir+rbs+slr+gfpf #2,3 (from last week)

Discussion

Quote of the Day

History is the only laboratory we have in which to test the consequences of thought. - Étienne Gilson

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