Lecture 18: Molecular Bio in Medicine Flashcards
Molecular biology has and will improve
human health
Molecular cloning is
Molecular cloning purposes
To obtain DNA of interest, one can use?
Polymerase Chain Reaction (PCR)
PCR amplifies?
Three steps of PCR
Advantages of PCR
Application of PCR
RT-PCR used to obtain?
PT-PCR enzyme
RT-PCR uses
RT-PCR protocol
RT-PCR can amplify…
Manipulate DNA for cloning uses
Restriction endonucleases (or restriction enzymes) function
Restriction enzymes
“Sticky” ends vs. blunt ends
Palindrome in dsDNA
Restriction enzymes can leave?
After PCR and endonucleases…
Gel electrophoresis
GE, molecules separate based on
Linear DNA separates by….
DNA staining with fluorescent dyes steps (4)
Polymorphism detection using?
RFLPs & example
Detection of RFLPs by PCR
DNA needs to be placed in
Plasmid/Vector
Plasmid/Vector uses
Plasmid DNA (cloning vector) contains three elements?
Molecular cloning
How to place target DNA into plasmid (2)
Making recombinant DNA main steps
B-lactamase
DNA Ligase in recombinant DNA (3 steps + note)
DNA sequencing: determine the exact order of nucleotides requires? What method?
To confirm the correct clone, what must happen?
dNTP
ddNTP
Dideoxynucleotide ddNTP function
DNA polymerase catalyzes?
Addition of a nucleotide mediated through 3’-OH group?
Incorporated dideoxynucleotide lack a?
Sequencing reaction mix contains all reagents including?
Sequencing protocol steps (4)
Each synthesized DNA has specific length
DNA length matched to
Fluorophore steps
Fluorescent color = ?, Migration in gel = ?
