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Team:Calgary/Notebook/Protocols/Process16
From 2011.igem.org
Notebook
Post-Regionals
Journal
- Defining Objectives
- Techniques Workshop
- Bioinformatics & qRT-PCR
- Sensory Element Fishing
- Electrochem - CPR Oxidation
- Electrochem - Reporter Gene
- Pseudomonas Tools
- Microalgae Tools
- NA Viability Assays
- May - July Group Journal
Protocols
Safety
Agarose Gel Electrophoresis
Reagents and Materials
- 1X TAE
- Graduated Cylinder
- 125 mL flask
- Agarose
- Gel Pouring Tray
- Tape
- Gel rig
- SYBR Safe
Protocol
- Measure out 100mL of buffer
- Transfer buffer to 125 mL flask
- Weigh out enough agarose to make a 1% gel (in our case 1.0 g of agarose was the right amount)
- Transfer agarose to 125mL flask
- Melt agarose in microwave until solution is almose boiling, stirring every 15-20 seconds (should be around 2 minutes)
- Allow agarose to cool (do not let it cool to the point where it is hard)
- Add 4 uL of SYBR Safe to the cooling agarose
- Assemble the gel pouring apparatus by inserting gate into slots.
- Allow gel to cool until flask can be handled comfortably.
- Place comb in the gel rig.
- Pour agarose into gel tray.
- Allow to solidify. While the gel is solidifying prepare the samples. Add your sample and 1 uL 10x Loading Dye, 4 uL of DNA and 5 uL of water
- Pour 1X TAE over gel so that gel is covered by a 3-5mm buffer
- Load samples into lane (Don't forget to load a 1kb+ ladder into one of the lanes)
- Hook electrodes to gel apparatus.
- Run the apparatus at 100V for 30 - 45 minutes (make sure to watch that the dye does not run off the gel)
- Visualize the gel and record the results
