Minnesota/5 August 2008

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|'''4. Run a gel:''' Using 1% agarose gel, check for correct amount/type of DNA bands.  
|'''4. Run a gel:''' Using 1% agarose gel, check for correct amount/type of DNA bands.  
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|'''5. Sequencing Rxns:''' Send in sequences. Follow the mixture below, which was sent in today:
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|'''5. Gel purification: The terminator band was cut out of the electrophoresis gel and purified out using QIAGEN's QuickSpin gel purification protocol. DNA used in ligation below.
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|'''6. Sequencing Rxns:''' Send in sequences. Follow the mixture below, which was sent in today:

Revision as of 20:47, 6 August 2008

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1. Plasmid Prep: Plasmid prep from the 2mL cultures made from picked colonies of (a) TetR/p22mnt + RFP + Term, (b) Lac/LAMBDA + GFP 1, (c) Lac/LAMBDA + GFP 2, (d) Tetpro + LAMBDAcI + term.
2. Spectrophotometry: Spec'ed the plasmid prep products to check for concentration of DNA.
3. Digest Rxn: Run a digest reaction on the plasmid prep products. Follow the chart below:


Parts 10x Buffer BSA H20 Insert DNA RE1 RE2 Total
L/L PCR 5.0uL 05uL 22.5uL 20.0uL 1.0uL, EcoRI 1.0uL, Pst1 50.0uL
TetR/p22mnt PCR 5.0uL 0.5uL 22.5uL 20.0uL 1.0uL, EcoRI 1.0uL, Pst1 50.0uL
BV 5.0uL 0.5uL 37.5uL 5.0uL 1.0uL, EcoRI 1.0uL, Pst1 50.0uL
Lac/LAMBDA + GFP 1 5.0uL 0.5uL 12.5uL 30.0uL 1.0uL, Spe1 1.0uL, Pst1 50.0uL
Term 5.0uL 0.5uL 37.5uL 5.0uL 1.0uL, Xba1 1.0uL, Pst1 50.0uL
L/L + GFP 2 5.0uL 0.5uL 7.5uL 35.0uL 1.0uL, Spe1 1.0uL, Pst1 50.0uL
4. Run a gel: Using 1% agarose gel, check for correct amount/type of DNA bands.
5. Gel purification: The terminator band was cut out of the electrophoresis gel and purified out using QIAGEN's QuickSpin gel purification protocol. DNA used in ligation below.
6. Sequencing Rxns: Send in sequences. Follow the mixture below, which was sent in today:


Parts H20 DNA primer
TetR/p22mnt + RFP + Term --- 8.8uL 3.2uL
TetR/p22mnt + RFP + Term --- 8.8uL 3.2uL
L/L + GFP 1A --- 8.8uL 3.2uL
L/L + GFP 1E --- 8.8uL 3.2uL
Tetpro + LAMBDAcI + TermA --- 8.8uL 3.2uL
Tetpro + LAMBDAcI + TermB --- 8.8uL 3.2uL
L/L + GFP 2A --- 8.8uL 3.2uL
L/L + GFP 2B --- 8.8uL 3.2uL