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5 Cards in this Set
- Front
- Back
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Gel electrophoresis (PAGE) |
-Separate via size, charge, and shape -Start at anode (+), end at cathode (-) -SDS - Denatures and gives proteins same (-) charge ratio, isolate size -Reducing - Reduces disulfide bonds -Native - Keeps protein in original shape, no denaturing or SDS |
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Blotting |
-Identify specific sequences after PAGE -Immobilize separated strands with UV light, use probe to identify sequence with reporter -Southern - DNA -Northern - RNA -Western - Protein (probe is a set of antibodies instead of a complementary strand) |
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Sanger sequencing |
-Determine sequence of DNA strand -Denature and add primer with dNTPs and ddNTPs -DNA poly extends with dNTPs -Randomly add ddNTPs, lack 3' -OH group, ending elongation -End with every possible unfinished DNA strand of varying lengths -PAGE separates, determine complementary sequence based off order of the ddNTP |
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PCR |
-Increase amount of DNA -Add DNA, primer, DNA poly, dNTPs (reverse transcriptase if needed) -Increase temp to denature DNA (need heat resistant DNA poly) -Lower temp, primer binds (annealing) -DNA poly elongates strand -Repeat, double strands after every round |
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ELISA |
-Determine if antibodies present against a certain antigen -Direct: Immobilize antigen and add antibody with reporter to see if it binds -Indirect: same as direct, but add second antibody with reporter to bind to 1st |
