Genetics 3

Question 1

molecular cloning=

Answer 1

process of growing large quantities of the DNA sequence of interest

Question 2

clone=

Answer 2

recombinant DNA molecule that has the same identical transferred segment DNA

Question 3

what do restriction enzymes do?

Answer 3

• they recognize specific double stranded sequence in DNA

- they cleave phosphodiester backbones of dsDNA at or near the recognition site

Question 4

what is a vector?

Answer 4

• a DNA molecule that can replicate autonomously in a host such as bacterial or yeast cells
• a plasmid

Question 5

what is a BAC

Answer 5

a bacterial artificial chromosome vectors

Question 6

human DNA fragment inserted into a vector molecule using what

Answer 6

DNA ligase

Question 7

what do plasmids consist of

Answer 7

circular, double stranded DNA molecules

Question 8

what is complementary DNA (cDNA) libraries

Answer 8

• contains only the exons thus is a direct representation of the coding
• can be derived from major mRNA transcripts that are selectively expressed

Question 9

what are the “ingredients for the perfect DNA”

Answer 9

• mRNA with a polyA tail
• reverse transcriptase
• cDNA strand
• vectors
• ligase

Question 10

what is nucleic acid hybridization used for?

Answer 10

-used to perform a “library screening” to ID clone carrying specific DNA insert of interest

Question 11

what is southern blotting used for?

Answer 11

standard method of analyzing specific fragments of DNA sequences cleaved by restriction enzymes

Question 12

major pit fall of southern blotting

Answer 12

does not identify mutations that are small like single base, insert, or deletes

Question 13

northern blotting involves

Answer 13

similar to southern blotting but involves the single stranded RNA

Question 14

what is PCR an alternative for?

Answer 14

alternative for cloning for generating essentially unlimited amounts of a sequence of interest

Question 15

how does PCR work? (basic)

Answer 15

• can selectively amplify a single molecule of DNA several billion fold in a few hours
• utilizes enzymatic application of a fragment of DNA

Question 16

what things are needed for PCR reaction

Answer 16

• fragment of DNA (target) between two oligonucleotide “primers”
• primers designed to be complementary to one strand of DNA molecule
• heat to denature double stranded fragments

Question 17

advantages of PCR

Answer 17

• can generate sufficient quantities of specific genes from DNA samples for the analysis of mutations
• sensitive, fast, less expensive, less sampling

Question 18

western blot is

Answer 18

technique in examining one or more proteins in a sample of cells or tissues

Question 19

what does western blot analyze specifically

Answer 19

-analysis of both normal and abnormal gene function through examination of protein encoded by the normal or mutant gene