AlanFung:Protocol/Agarose Gel Synthesis: Difference between revisions
Jump to navigation
Jump to search
>Alan6017518 |
>Alan6017518 |
||
(3 intermediate revisions by the same user not shown) | |||
Line 26: | Line 26: | ||
==Protocol== | ==Protocol== | ||
*Determine the concentration of Agarose to be used | |||
(1 Tray) | |||
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 | ||
*Weigh out the required amount of Agarose with a scale | |||
* | *Measure the required volume of TBE buffer using a graduated cylinder | ||
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 | ||
by swirling the flask and microwave for 30s | *Cast gel into tray with the required comb set in the tray | ||
powder are dissolved and the solution appears to be clear | *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