recombinant DNA technology

Question 1

recombinant DNA (rDNA)

Answer 1

DNA that’s been artificially created by combining genetic material from multiple sources

Question 2

how is recombinant DNA achieved

Answer 2

molecular cloning

Question 3

steps to clone rDNA

Answer 3

-isolate DNA segment to be cloned
-select a cloning vector
-join DNA fragments together
-clone rDNA in host organism

Question 4

getting an rDNA in host organism

Answer 4

-transformation
-transfection
-transduction

Question 5

transformation

Answer 5

uptake of exogenous DNA from environment by a bacterial cell (cells must be competent)

Question 6

transfection

Answer 6

forced introduction of a small molecule into eukaryotic cells
-chemical
-lipid based
-electroporation

Question 7

transduction

Answer 7

introduction of exogenous genetic material into cells by a virus

Question 8

3 steps

Answer 8

-denature (raise temp to 95)
-anneal primers (lower temp to 60)
-extend (polymerase)(raise temp to 72)

Question 9

probe

Answer 9

(primer)
DNA or RNA with a base sequence complimentary to a sequence in the gene of interest

Question 10

southern blot

Answer 10

DNA

Question 11

northern blot

Answer 11

RNA

Question 12

FISH

Answer 12

fluorescence in situ hybridization
-probes used to determine the chromosomal location and to visualize a genes expression while its in a cell

Question 13

taq-probe

Answer 13

used in qRT-PCR to detect specific DNA sequences with high accuracy

Question 14

dideoxy sequencing

Answer 14

uses dideoxynucleotides (ddNTPs) to determine the sequence of bases in a DNA fragment
-uses a different fluorescent dye to distinguish the four bases (A,T,C,G)