Team:Lethbridge/Notebook/Lab Work/June
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Analyzed results on a 1% agarose gel. Load order as follows:<br> | Analyzed results on a 1% agarose gel. Load order as follows:<br> | ||
<table><table border ="3"> | <table><table border ="3"> | ||
- | <tr><td><b>Lane</b></td><td><b>Sample</b></td><td><b>Volume<br>Sample (µL)</b></td><td>Volume Loading<br>Dye (µL)</b></td></tr> | + | <tr><td><b>Lane</b></td><td><b>Sample</b></td><td><b>Volume<br>Sample (µL)</b></td><td><b>Volume Loading<br>Dye (µL)</b></td></tr><tr><td>1</td><td>Restricted RBS-xylE</td><td>10</td><td>2</td></tr> |
- | <tr><td>1</td><td>Restricted RBS-xylE</td><td>10</td><td>2</td></tr> | + | |
<tr><td>1</td><td>Unestricted RBS-xylE<sup>†</sup></td><td>1</td><td>2</td></tr> | <tr><td>1</td><td>Unestricted RBS-xylE<sup>†</sup></td><td>1</td><td>2</td></tr> | ||
<tr><td>1</td><td>1kb Ladder<sup>†</sup><sup>†</sup></td><td>2</td><td>2</td></tr></table> | <tr><td>1</td><td>1kb Ladder<sup>†</sup><sup>†</sup></td><td>2</td><td>2</td></tr></table> |
Revision as of 00:30, 11 June 2010
Contents |
June 2010
June 1/2010
JV quantified the amount of DNA in gels run to date using ImageJ software. Results to be posted in working plasmids box.
Objective: Transform plasmids into DH5α
Method: Follow competent cell transformation protocol to transform the following:
From our ligations:
- pLacI-sRBS-Lumazine-dT
- pLacI-sRBS-Lumazine-dT
- mms6 (A6)
- mms6 (B6)
- xylE (C4)
- xylE (B4)
From the 2010 Parts Distribution:
- ECFP (Bba_E0020)
- EYFP (Bba_E0030)
- BglII Endonuclease (Bba_K112106)
June 2/2010
(In Lab: JV)
Objective: Isolate plasmid DNA of RBS-xylE (BBa_J33204) from DH5α cells and confirm results.
Method: "Mini-prep" the plasmid DNA using boiling lysis miniprep. Then restrict the DNA once and run on a 1% agarose gel (TAE).
Restriction Reaction
Ingredient | Volume(µL) |
MilliQ H20 Water | 15.75 |
Orange Buffer (10x) | 2 |
pDNA (rbs-xylE) | 2 |
EcoRI | 0.25 |
Unrestricted Control
Ingredient | Volume(µL) |
MilliQ H20 Water | 16 |
Orange Buffer (10x) | 2 |
pDNA (rbs-xylE) | 2 |
DNA was restricted for 80 minutes at 37oC.
Analyzed results on a 1% agarose gel. Load order as follows:
Lane | Sample | Volume Sample (µL) | Volume Loading Dye (µL) |
1 | Restricted RBS-xylE | 10 | 2 |
1 | Unestricted RBS-xylE† | 1 | 2 |
1 | 1kb Ladder†† | 2 | 2 |
† Added 9µL MilliQ H2O
†† Added 8µL MilliQ H2O