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BISC219/F12: RNAi Lab 7: Difference between revisions
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→Agarose Gel Electrophoresis
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#Add 5 μL of loading dye to each PCR product. You will find a precast 1% agarose gel with Sybr-Safe in 1x TAE buffer (prepared by our lab specialist) that is just for two groups' 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> | #Add 5 μL of loading dye to each PCR product. You will find a precast 1% agarose gel with Sybr-Safe in 1x TAE buffer (prepared by our lab specialist) that is just for two groups' 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, A, B, C. <BR> | #Load 15μL of each PCR product into separate wells in the gel. Make sure they are loaded in alphabetical order left to right, A, B, C. <BR> | ||
#Load 5 μL of the 100 base pair DNA ladder in the well on the far right. Our standards ladder is product number NEB323-2S from New England Biologicals. The manufacturer supplies the following information about | #Load 5 μL of the 100 base pair DNA ladder in the well on the far right. | ||
#Run the gel for ~ 20min. at ~120 volts. <BR> | |||
Our standards ladder is product number NEB323-2S from New England Biologicals. The manufacturer supplies the following information about the standards ladder.<BR> | |||
[[Image:NEB323-S1.jpg]] | [[Image:NEB323-S1.jpg]] | ||
<BR> | <BR> | ||
=='''Capturing Digital Images of Nucleic Acid Gels Stained with SYBR Safe Using the BioRad Imaging System in L308''' == | =='''Capturing Digital Images of Nucleic Acid Gels Stained with SYBR Safe Using the BioRad Imaging System in L308''' == | ||
