©Themoon EXAM SOLUTIONS
25/11/2024 09:27AM
BIOC 301 UBC Final Exam and Answers
What type of matrix can be used for Ion Exchange Chromatography? - Answers✓✓Insoluble
matter (beads) containing covalently bound charged residues
Anion exchange chromatography - Answers✓✓Negative proteins stick to positive beads, only
positive proteins go through
Cation Exchange Chromatography - Answers✓✓Positive proteins stick to negative beads, only
negative proteins go through
How do we choose a matrix based on the properties of the protein to be isolated? -
Answers✓✓Based on protein net charge, pI, pH and accessibility of charged residues
How do we remove contaminants from the column? - Answers✓✓Wash buffer
How do we remove the protein from te column? (two ways) - Answers✓✓1) Increasing ionic
strength of the buffer
2) Changing pH of the buffer
Variables in crystallization - Answers✓✓1) Amount of starting material
2) Level of purity of the protein
3) Solvent (pH and concentration)
4) Precipitant (type and concentration)
, ©Themoon EXAM SOLUTIONS
25/11/2024 09:27AM
Why is crystallization desirable? - Answers✓✓Allows to perform X-ray crystallography
How to test if crystals are proteins? (4 ways) - Answers✓✓1) Dehydration (dried salt crystal
remains intact but protein one does not)
2) Crystals tend to be birefringent
3) Dye absorption (salt crystals don't absorb dye)
4) Run SDS-PAGE
Solutions used in lysis protocol - Answers✓✓Lysis solution, Neutralization solution, Wash
solution, and Elution Buffer
Lysis solution (alkaline plasmid isolation) - Answers✓✓SDS+Strong base
Disrupt cells' membranes
Neutralization solution (plasmid isolation) - Answers✓✓Precipitates (denatures) genomic DNA
Wash Solution (plasmid isolation) - Answers✓✓Removes salt residues on membrane
Elution Buffer (plasmid isolation) - Answers✓✓Dissolves DNA in slightly basic buffer
How is plasmid DNA separated from genomic DNA? - Answers✓✓Denatured (after lysis
solution) genomic DNA can't reanneal as fast as plasmid DNA, thus it gets precipitated.
Common features of usefuk plasmids (3) - Answers✓✓1) Origin of replication
2) A drug-resistance gene
25/11/2024 09:27AM
BIOC 301 UBC Final Exam and Answers
What type of matrix can be used for Ion Exchange Chromatography? - Answers✓✓Insoluble
matter (beads) containing covalently bound charged residues
Anion exchange chromatography - Answers✓✓Negative proteins stick to positive beads, only
positive proteins go through
Cation Exchange Chromatography - Answers✓✓Positive proteins stick to negative beads, only
negative proteins go through
How do we choose a matrix based on the properties of the protein to be isolated? -
Answers✓✓Based on protein net charge, pI, pH and accessibility of charged residues
How do we remove contaminants from the column? - Answers✓✓Wash buffer
How do we remove the protein from te column? (two ways) - Answers✓✓1) Increasing ionic
strength of the buffer
2) Changing pH of the buffer
Variables in crystallization - Answers✓✓1) Amount of starting material
2) Level of purity of the protein
3) Solvent (pH and concentration)
4) Precipitant (type and concentration)
, ©Themoon EXAM SOLUTIONS
25/11/2024 09:27AM
Why is crystallization desirable? - Answers✓✓Allows to perform X-ray crystallography
How to test if crystals are proteins? (4 ways) - Answers✓✓1) Dehydration (dried salt crystal
remains intact but protein one does not)
2) Crystals tend to be birefringent
3) Dye absorption (salt crystals don't absorb dye)
4) Run SDS-PAGE
Solutions used in lysis protocol - Answers✓✓Lysis solution, Neutralization solution, Wash
solution, and Elution Buffer
Lysis solution (alkaline plasmid isolation) - Answers✓✓SDS+Strong base
Disrupt cells' membranes
Neutralization solution (plasmid isolation) - Answers✓✓Precipitates (denatures) genomic DNA
Wash Solution (plasmid isolation) - Answers✓✓Removes salt residues on membrane
Elution Buffer (plasmid isolation) - Answers✓✓Dissolves DNA in slightly basic buffer
How is plasmid DNA separated from genomic DNA? - Answers✓✓Denatured (after lysis
solution) genomic DNA can't reanneal as fast as plasmid DNA, thus it gets precipitated.
Common features of usefuk plasmids (3) - Answers✓✓1) Origin of replication
2) A drug-resistance gene
