Team:StanfordBrownSpelman/Lab Techniques4

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<br />Gel should be placed on scanner face-up. That is, the wells should be oriented up, the same way the gel is oriented in the gel box
<br />Gel should be placed on scanner face-up. That is, the wells should be oriented up, the same way the gel is oriented in the gel box
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<br /><b>Gel Extraction & Cleanup</b>
<br /><b>Gel Extraction & Cleanup</b>
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<br />Make sure to place gel on transilluminator face down (wells toward the glass). Remove as much excess gel matrix as possible without overexposing DNA to UV.
<br />Make sure to place gel on transilluminator face down (wells toward the glass). Remove as much excess gel matrix as possible without overexposing DNA to UV.
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Latest revision as of 03:11, 16 October 2014

Stanford–Brown–Spelman iGEM 2014 — Amberless Hell Cell

Verification
Gel Casting:

0.75% agarose (if DNA>1000bp)
40mL 1x TAE
0.3 g agarose
1 aliquot (~5μl) gel red

Add dry agarose to clean bottle (small enough to fit in microwave)
Add 40mL 1x TAE buffer
Microwave with cap on but loose, swish periodically, until solution is clear and smooth
Pipette in gel red, directly into solution (heat stable so don’t worry about the temperature)
Pour into gel tray, making sure that tray is oriented and tightly inserted such that leaks will not occur, and that the gel is level

Gel Loading & Running

Lane 1 should be ladder; use 1kb ladder or 100bp ladder depending on the size of your DNA samples
Digests can require more (~1.5x) than the usual amount of loading dye

Gel Imaging (using Typhoon scanner)
Always scan a gel immediately after running
Make sure the scanner area is clean; wipe ONLY with 70% ethanol (or DI) and kimtech wipes
Gel should be placed on scanner face-up. That is, the wells should be oriented up, the same way the gel is oriented in the gel box

Gel Extraction & Cleanup

Make sure to place gel on transilluminator face down (wells toward the glass). Remove as much excess gel matrix as possible without overexposing DNA to UV.

For cleanup, follow protocol for using the Wizard PCR Cleanup Kit, found in PCR section.
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