MCB3020 Exam 3 Questions and
Answers 100% Correct
Reverse transcriptase process - ANSWER - 1. poly-T primer to single-strand RNA
2. reverse transcriptase to produce DNA-RNA hybrid
3. enzyme RNaseH to cut RNA and regenerate RNA primers
4, DNA polymerase and ligase synthesizes new complementary DNA strand
cloning vector - ANSWER - allows for transfer of gene of interest to a host organism; allows for
the making of recombinant DNA
good cloning vector qualities - ANSWER - origin of replication
selectable marker (i.e. antibiotic resistance gene)
unique restriction sites that allow the insertion of gene of interest
types of cloning vectors - ANSWER - plasmid (carry 20k base pairs)
bacteriophage (carry 9k-25k base pairs)
cosmid (carry 30k-47k base pairs)
artificial chromosomes (carry up to 1 million base pairs)
primary metabolite - ANSWER - essential for proper growth, its removal significantly impacts
survival
,secondary metabolite - ANSWER - not directly involved in growth; its removal doesn't
significantly impact survival;
productions is usually triggered by exogenous events
protoplast fustion - ANSWER - cells without cell walls are put together in solution, then shocked
electrically to fuse them all together
chemostat - ANSWER - bioreactor with a constant volume caused by a constant inflow of fresh
medium and output of culture liquid
PCR temperatures - ANSWER - 90 - separates DNA strands
50 - primers attach
70 - nucleotides are added on
types of polymers for gel electrophoresis - ANSWER - agrose
, polyacrylimide
ethidium bromide - ANSWER - stains DNA, makes it visible under UV
RFLP Analysis procedure - ANSWER - 1. isolate gene of interest
2. PCR
3. cut DNA, making restriction fragments
4. gel electrophoresis to separate according to size
5. computers to analyze
ribotyping - ANSWER - compares rRNA to find similarities/differences across domains
southern blotting - ANSWER - 1. cut DNA to make a lot of fragments
2. separate DNA by size via gel electrophoresis
3. place sheet of nitocellulose paper on top of gel
4. place paper in heat-sealed bag with radioactive probe
5. wash filter and place on x-ray film to make autoradiograph
western blotting - ANSWER - detects proteins
northern blotting - ANSWER - detects RNA
making of a genomic library - ANSWER - 1. extract DNA
2. cut into fragments of specific size
3. insert the fragments into a vector
4. insert vector to host organism
5. DNA can now be amplified/retrieved
site-directed mutagenesis - ANSWER - 1. identify sequence
2. complementary DNA sequence in lab with the mutation
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