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)
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
3. insert the sequence into target cell
4. target cell swaps target sequence with complementary sequence with mutation
protoplast fustion - answer✔✔cells without cell walls are put together in solution, then shocked
electrically to fuse them all together
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
biosurfactants - answer✔✔emulsify another substance;
useful in oil recovery, dispersing oil spills, and bioremediation
bioconversion - answer✔✔live organisms facilitate chemical rxns
i.e. microorganisms as biocatalysts; immobilized cells work better cuz they can be reused
biosensors - answer✔✔microorganisms, enzymes, or organelles detect the presence/absence of a
substance and converts products of the rxn to electrical currents
horizontal gene transfer - answer✔✔creates recombinant DNA
3 types: conjugation, transformation, transduction
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