AlanFung:Protocol/Agarose Gel Synthesis: Difference between revisions
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DNA Size 5 - 10 kb 0.2 - 1 kb | DNA Size 5 - 10 kb 0.2 - 1 kb | ||
Concentration of Agarose 0.7 % 2 % | Concentration of Agarose 0.7 % 2 % | ||
Amount of Agarose 0.7 g 2 g | Amount of Agarose 0.7 g 2 g | ||
Amount of 0.5X TBE Buffer 100 mL 100 mL | Amount of 0.5X TBE Buffer 100 mL 100 mL | ||
Amount of SYBR Safe 8uL 8uL | Amount of SYBR Safe 8uL 8uL | ||
Note: Scale up or down accordingly | Note: Scale up or down accordingly |
Revision as of 22:39, 22 June 2009
Agarose_Gel_Synthesis_Protocol PDF
Agarose Gel Synthesis Protocol
Overview
*Determine the concentration of agarose to be used *Measurement of agarose gel and TBE buffer *Mix and Heat *Cool Down *Gel Staining *Casting *Storage
Samples & Material
*Erlenmeyer Flask (250mL) *Graduated Cylinder *Mircowave *Gel tray 13 x 12.2cm (2 standard gel trays) *Gel Tray (4 micro gel trays) *Combs for micro gels (6/8 teeth) *Standard combs (13/26 teeth) *Gel casting stand for micro gels *Gel casting stand *0.5X TBE Buffer *Ultra Pure Agarose -Invitrogen Cat:15510-027 *SYBR Safe DNA gel stain10,000X concentration in DMSO -Invitrogen Cat:S33102
Protocol
1-Determine the concentration of Agarose to be used
DNA Size 5 - 10 kb 0.2 - 1 kb Concentration of Agarose 0.7 % 2 % Amount of Agarose 0.7 g 2 g Amount of 0.5X TBE Buffer 100 mL 100 mL Amount of SYBR Safe 8uL 8uL
Note: Scale up or down accordingly
- 2-Weigh out the required amount of Agarose with a scale and measure the required volume of
TBE buffer using a graduated cylinder
- Caution: Hot Agarose solution should be handled very carefully.
- 3-Transfer the Agarose powder into a 250mL Erlenmeyer flask and add in the TBE buffer, mix
by swirling the flask and microwave for 30s, repeat mixing and heating step until all gel powder are dissolved and the solution appears to be clear.