IGEM:PennState/2006/dnagels

'' =AGAROSE (DNA) GEL PROTOCOL=

Media/Reagents

 * 1X TAE or 1X TBE
 * Agarose (powder)

Gel

 * 1) Make [x] % agarose gel (%= weight/volume) according to size DNA you want to separate (higher MW bands are better resolved by lower percentage gels, e.g. 0.6%, while lower MW bands are best resolved by higher percentage gels, e.g. 1.4%). For most applications, 1% gel is appropriate, so weigh 1g agarose, and place in a 250 mL erlenmeyer flask (or appropriately large glassware so that the solution won’t boil over).
 * 2) To it add 100 mL 1X TAE or 1X TBE (TAE is used when DNA will be extracted from gel, TBE is used for diagnostic purposes).
 * 3) Swirl to mix.
 * 4) Cover flask w/ Reynolds wrap & microwave for approx. 45 sec or until solution begins to boil.
 * 5) Cool flask until it is hold-able. Note: >55C may warp the tray.
 * 6) Pour into gel box and allow to solidify, don't forget the combs!

Loading Gel:

 * 1) Add x10 SYBR green to DNA and incubate for 15 in dark
 * 2) Add x5 loading buffer
 * 3) Add 2 μL loading buffer to each sample and load on gel
 * 4) Add samples to wells being care not to spill over into adjacent wells
 * 5) Run at 100 V until bands migrate approx. 2/3 down gel. (takes around 45 min)