AlanFung:Protocol/Agarose Gel Synthesis: Difference between revisions
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[[Media:Agarose Gel_Synthesis_Protocol.pdf|Agarose_Gel_Synthesis_Protocol PDF]] | [[Media:Agarose Gel_Synthesis_Protocol.pdf|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== | |||
*Determine the concentration of Agarose to be used | |||
(1 Tray) | |||
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 | |||
*Weigh out the required amount of Agarose with a scale | |||
*Measure the required volume of TBE buffer using a graduated cylinder | |||
Caution: Hot Agarose solution should be handled very carefully. | |||
*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 | |||
*Let cool for a while and add in the required amount of SYBR Safe, swirl flask to mix the dye | |||
*Cast gel into tray with the required comb set in the tray | |||
*Let cool |
Latest revision as of 22:51, 22 June 2009
Agarose_Gel_Synthesis_Protocol PDF
Agarose Gel Synthesis Protocol[edit]
Overview[edit]
*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[edit]
*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[edit]
- Determine the concentration of Agarose to be used
(1 Tray) 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
- Weigh out the required amount of Agarose with a scale
- Measure the required volume of TBE buffer using a graduated cylinder
Caution: Hot Agarose solution should be handled very carefully.
- 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
- Let cool for a while and add in the required amount of SYBR Safe, swirl flask to mix the dye
- Cast gel into tray with the required comb set in the tray
- Let cool