Genetics 3 Flashcards
molecular cloning=
process of growing large quantities of the DNA sequence of interest
clone=
recombinant DNA molecule that has the same identical transferred segment DNA
what do restriction enzymes do?
- they recognize specific double stranded sequence in DNA
- they cleave phosphodiester backbones of dsDNA at or near the recognition site
what is a vector?
- a DNA molecule that can replicate autonomously in a host such as bacterial or yeast cells
- a plasmid
what is a BAC
a bacterial artificial chromosome vectors
human DNA fragment inserted into a vector molecule using what
DNA ligase
what do plasmids consist of
circular, double stranded DNA molecules
what is complementary DNA (cDNA) libraries
- contains only the exons thus is a direct representation of the coding
- can be derived from major mRNA transcripts that are selectively expressed
what are the “ingredients for the perfect DNA”
- mRNA with a polyA tail
- reverse transcriptase
- cDNA strand
- vectors
- ligase
what is nucleic acid hybridization used for?
-used to perform a “library screening” to ID clone carrying specific DNA insert of interest
what is southern blotting used for?
standard method of analyzing specific fragments of DNA sequences cleaved by restriction enzymes
major pit fall of southern blotting
does not identify mutations that are small like single base, insert, or deletes
northern blotting involves
similar to southern blotting but involves the single stranded RNA
what is PCR an alternative for?
alternative for cloning for generating essentially unlimited amounts of a sequence of interest
how does PCR work? (basic)
- can selectively amplify a single molecule of DNA several billion fold in a few hours
- utilizes enzymatic application of a fragment of DNA
