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BISC219/F12: RNAi Lab 7: Difference between revisions
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→Agarose Gel Electrophoresis
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=='''Agarose Gel Electrophoresis'''== | =='''Agarose Gel Electrophoresis'''== | ||
After the PCR reactions | After the PCR reactions have, we hope, made millions of copies of ''lys-2'' you will take a sample of the pcr product and run a gel to analyze the results of the amplification (the search for your gene). <BR> | ||
Add 5 μL of loading dye to each PCR | Add 5 μL of loading dye to each PCR product.<BR> | ||
You will find a precast 1% agarose gel with Sybr-Safe in 1x TAE buffer that is just for your group's use. Make sure that the wells of the gel are closest to the black (negative) electrode and that the gel apparatus has plenty of buffer. Draw a template in your lab notebook so you know which colony is to be put in each lane (1-7 left to right) and which lane contains your DNA ladder. Make sure you get a copy of the DNA ladder key.<BR> | You will find a precast 1% agarose gel with Sybr-Safe in 1x TAE buffer (prepared by our lab specialist) that is just for your group's use. Make sure that the wells of the gel are closest to the black (negative) electrode and that the gel apparatus has plenty of buffer. Draw a template in your lab notebook so you know which colony is to be put in each lane (1-7 left to right) and which lane contains your DNA ladder. Make sure you get a copy of the DNA ladder key.<BR> | ||
Load 15μL of each PCR product into separate wells in the gel. Make sure they are loaded in alphabetical order left to right. <BR> | Load 15μL of each PCR product into separate wells in the gel. Make sure they are loaded in alphabetical order left to right. <BR> | ||
Load 5 μL of the 100 base pair DNA ladder in the well on the far right.<BR> | Load 5 μL of the 100 base pair DNA ladder in the well on the far right.<BR> | ||
