Molecular Methods I: DNA Cloning and Sequencing Flashcards
What class of enzyme found in bacteria cleaves double-stranded DNA at highly specific sequences?
Restriction Endonucleses(Enzymes)
What is the general pattern of the sequence of restriction site?
Palindromic
How are restriction enzymes named?
1st letter- bacteria genus; 2nd, 3rd- species; 4th-type or strain; Number- order of discovery
What molecule is left on the 3’ end of the DNA following cleavage? The 5’ end?
3’- hydroxyl group; 5’- phosphate group
What are the two types of ends that can be generated from restriction enzyme digestion?
Blunt and Sticky ends
What enzyme covalently attaches hybridized DNA together?
DNA Ligase
What is the formula for calculating how often a sequence of (n) bases will occur?
every 4^n bps
What are the three essential properties of vectors?
Capable of autonomous replication, contains at least one unique restriction site, has at least one selectable marker
What control is used to ensure only transformed bacteria survive?
Selectable markers (Ampicillin, Neomycin)
What genetic feature of plasmids allows them to replicate independent of the host?
Origins of replication
What is the process by which recombinant DNA is introduced into bacterial hosts?
Transformation
What is a collection of cloned restriction fragment of the DNA of an organism?
DNA Library
What are the two basic forms of DNA Libraries?
Genomic and Complementary DNA Libraries
Which DNA library contains DNA derived solely from expressed genes?
Complementary DNA Libraries
What unique enzyme is used in the creation of cDNA?
Reverse Transcriptase
