Team:Paris Bettencourt/Protocols/Gel Electrophoresis

Ferment It Yourself

iGEM Paris-Bettencourt 2O15

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Vitamin A	Vitamin B2	Vitamin B12
Phytase	Riboswitch	Differentiation on E. coli
Differentiation on S. cerevisiae	Manufacturing	Idli and Micro-organisms

Gel-Electrophoresis protocol Preparation of the Gel (50mL) Mix 0.5 grams of agarose with 50mL 0.5% TAE solution Heat the Erlenmeyer in a microwave until the liquid becomes totally transparent. Let it rest until you can take the Erlenmeyer in your hand without pain for 10 seconds. Add 5uL of SYBR® Safe (10000X) and mix gently until the liquid takes and uniform reddish color. Pour the liquid in a gel mold with the desired number of well and wait 20-30 min (or until the gel is solidified). Put the gel inside the electrophoresis machine and pour some TAE 0.5% until the gel is totally submerged. Preparation of the samples Mix 5uL of each sample with 1uL of 6X loading dye in microcentrifuge tubes/ on a piece of parafilm / in PCR tubes. Pipette 5-6uL of each sample inside a different well. Pipette 5uL of the appropriate ladder in one of the wells. Close the machine and launch the migration with the desired voltage After the desired migration time turn of the machine, remove the lid and put the gel inside the Blue light revelator.