BIO 204 Exam study guide with complete solutions | Rated A+
1. DNA polymerase: strings DNA tgt using single-stranded DNA as a template + requires primer
2. Taq polymerase: A heat-stable form of DNA polymerase
3. Buffer: maintain pH + ionic composition
4. Mg^2+: shield DNA polymerase from negatively charged phosphates on DNA backbone
5. dNTPs: substrate for DNA polymerase, 2/3 phosphate groups hydrolyzed for E for polymerase rxns
6. PCR thermal cycler: 1) Denature - 2x strand-> 1x strand (94C)
2) Anneal - primers + taq attach (55-65C)
3) Elongation - taq adds nucleotides (72C)
7. Equation for number of DNA fragments produced in PCR: X(2^n)
x= # of DNA molecules you start w
n= # of complete PCR cycles
8. Edward's buffer: Tris, SDS, pH 8, NaCl, EDTA
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, 9. Tris buffer: maintains pH and stabilizes DNA macromolecules
10. SDS: Anionic detergent that denatures proteins
11. EDTA: binds cations like Mg^2+ and Mn^2+ that are req for DNAses activity
12. NaCl: Na+ interacts w DNA phosphate backbone but needs alc (IPA) to first disrupt interactions btwn H2O and
DNA phosphate backbone
13. DMSO: improves amplification, disrupts secondary structures, stabilizes DNA, + enhances prime annealing
14. TE buffer: stabilizes DNA during storage
15. RNAse A: breaks down RNA into RNA fragments (bcs tm RNA can cause enzymes to not work well)
16. DNA extraction steps: 1) homogenize leaf in edwards buffer (break cell walls)
2) heat sample to 100C (denature proteins)
3) spin down homogenized sample (separate cellular debris from DNA)
4) discard pellet (keep supernatant w DNA)
5) add IPA (precipitate DNA)
6) spin down precipitated DNA (pellet DNA)
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1. DNA polymerase: strings DNA tgt using single-stranded DNA as a template + requires primer
2. Taq polymerase: A heat-stable form of DNA polymerase
3. Buffer: maintain pH + ionic composition
4. Mg^2+: shield DNA polymerase from negatively charged phosphates on DNA backbone
5. dNTPs: substrate for DNA polymerase, 2/3 phosphate groups hydrolyzed for E for polymerase rxns
6. PCR thermal cycler: 1) Denature - 2x strand-> 1x strand (94C)
2) Anneal - primers + taq attach (55-65C)
3) Elongation - taq adds nucleotides (72C)
7. Equation for number of DNA fragments produced in PCR: X(2^n)
x= # of DNA molecules you start w
n= # of complete PCR cycles
8. Edward's buffer: Tris, SDS, pH 8, NaCl, EDTA
1/4
, 9. Tris buffer: maintains pH and stabilizes DNA macromolecules
10. SDS: Anionic detergent that denatures proteins
11. EDTA: binds cations like Mg^2+ and Mn^2+ that are req for DNAses activity
12. NaCl: Na+ interacts w DNA phosphate backbone but needs alc (IPA) to first disrupt interactions btwn H2O and
DNA phosphate backbone
13. DMSO: improves amplification, disrupts secondary structures, stabilizes DNA, + enhances prime annealing
14. TE buffer: stabilizes DNA during storage
15. RNAse A: breaks down RNA into RNA fragments (bcs tm RNA can cause enzymes to not work well)
16. DNA extraction steps: 1) homogenize leaf in edwards buffer (break cell walls)
2) heat sample to 100C (denature proteins)
3) spin down homogenized sample (separate cellular debris from DNA)
4) discard pellet (keep supernatant w DNA)
5) add IPA (precipitate DNA)
6) spin down precipitated DNA (pellet DNA)
2/4
