Exam (elaborations)
FS-422 SEROLOGY AND DNA EXTRACTION
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FS-422 SEROLOGY AND DNA EXTRACTION
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FS-422 SEROLOGY AND DNA EXTRACTIONContamination Management - Answers-Wear appropriate PPE, standard lab practices
(gloves, tip exchange), use reagent blank, start with Q sample first.
Presumptive Testing (Blood) - Answers-Kastle Meyer
Confirmatory Testing (Blood) - Answers-RSID Blood
Kastle Meyer - Answers-color test for hemoglobin with phenolphthalein to produce a
deep pink (cannot distinguish b/w human/animal, plants can produce false pos, fecal
matter)
Organic DNA extraction - Answers-involves lysing open cells, treating with phenol
chloroform, and then treating aqueous layer with ethanol.
Chelex - Answers-Add water, add Chelex resin, boil and centrifuge, remove ALL Chelex
(PCR inhibitor)
Solid-Phase Extraction - Answers-lysis buffer (containing salt), transferred to spin
column (silica base), wash buffer (with high salt), eluted (low to no salt)
All DNA Extractions Must: - Answers-disrupt cells/tissues, inactivate DNA, denature
proteins, separate nucleic acids
FTA Paper - Answers-Stores blood samples, can be punched, washed, and instantly
used in PCR.
DTT - Answers-breaks disulfide bonds in presence of strong proteins (used in semen
and hair)
SDS - Answers-detergent used to solubilize cell membranes
Proteinase K - Answers-enzyme that degrades proteins
EDTA - Answers-(in buffer solution) used to disrupt cell membranes and inhibit
nucleases (decalcifies bones)
RSID Human Blood - Answers-Detects for the presence of human glycophorin A.
(specific and abundant in human red blood cells)
ABACard Hematrace - Answers-Works like RSID Blood but detects for hemoglobin
Presumptive Test (Semen) - Answers-Acid-Phosphatase
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