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Microbio Lab 7
Agarose Gel Electrophoresis and Electroporation
Question | Answer |
---|---|
components of DNA loading dye | glycerol, zylene cyanol and bromophenol blue. purpose = visualize gel progress and determine rate of travel |
glycerol | makes the sample heavier so it doesn't flow out of well |
xylene cyanol and bromophenol blue | dyes for sample |
which runs faster? small or large pieces | smaller |
Gel Star/SYBR Green | dye binds to DNA and fluoresces under UV light -visualizes and stains DNA fragments |
3 possible forms of undigested plasmid | supercoiled, nicked, and concatimerized |
Mss1 | restriction enzyme that linearizes plasmid DNA |
if loud popping is heard during electroporation | there is salt and poor transformation and bubbles |
electroporation | method by which yeast DNA is transformed, cells are shocked into uptake of naked DNA. >90% of cells dies |
why are cells allowed to recover? | to allow for expression (transcription and translation) of gene |
mat alpha | signals secretion of HRP out of cell |
sorbitol | osmotic stabilizer that prevents cell lysis. |