Basic procedures for bacteria culture(3)
时间:2023-08-16 23:12 来源:网络整理 作者:墨客科技 点击:次
1. Prepare an agarose gel, according to recipes listed below, by combining the agarose (low gel temperature agarose may also be used) and water in a 500 ml Ehrlenmeyer flask, and heating in a microwave for 2-4 minutes until the agarose is dissolved. 0.7% 1.0% 2.0% agarose 1.05 g 1.5 g 3.0 g 20X TAE 7.5 ml 7.5 ml 7.5 ml ddH2O 142.5 ml 142.5 ml 142.5 ml EtBr (5 mg/ml) 25 ul 25 ul 25 ul total vol 150 ml 150 ml 150 mlGenetic technology grade (800669) or low gel temperature (800259) agarose from Schwarz/Mann Biotech. 2. Add 20X TAE and ethidium bromide (EtBr), swirl to mix, and pour the gel onto a taped plate with casting combs in place. Allow 20-30 minutes for solidification. 3. Carefully remove the tape and the gel casting combs and place the gel in a horizontal electrophoresis apparatus. Add 1X TAE electrophoresis buffer to the reservoirs until the buffer just covers the agarose gel. 4. Add at least one-tenth volume of 10X agarose gel loading dye to each DNA sample, mix, and load into the wells. Electrophorese the gel at 150-200 mA until the required separation has been achieved, usually 0.5-1 hour (100-120 mA for low gel temperature agarose), and cool the gel during electrophoresis with a fan. Visualize the DNA fragments on a long wave UV light box and photograph with a Polaroid camera. (责任编辑：admin) |
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