Team:BCCS-Bristol/Protocols-Agarose Gel Electrophoresis

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==Agarose Gel Electrophoresis==

===Gel preparation:===

#	For a thick gel in a small chamber, boil 50 ml 0.5x TBE in the microwave (for a thinner gel to insert only 5 µl sample in each well 40 ml are sufficient)
#	Cool the solution until you can touch it with your hands for a longer time
#	Add 0.5 µl ethidium bromide per 10 ml TBE and mix while avoiding air bubbles
#	Pour the gel into a chamber with a comb that you prepared before
#	Let the gel cool down and become solid (15-20 min)

===Sample preparation: ===

#	Add 10 % “Sample Loading Buffer” (BIORAD) to the sample
#	Mix gently and spin shortly down

===Loading the gel: ===

#	Remove the comb and put the gel with the slide into a chamber (the wells need to be on the end with the black pole!!!)
#	Fill the chamber with 0.5x TBE until the gel is covered
#	Use 5 µl HyperLadderI (BIOLINE)
#	For colony PCR, put 5 µl of each reaction in the well
#	Close the chamber with the lid (black pole to black=negatively charged and red to red=positively charged…)
#	Run the gel with 80-100 V


The Sample Loading Buffer contains two dyes: Bromophenol blue runs at ~300 bp and Xylene cyanol FF runs at ~4 kb.

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