Chapter 5 - Exploring Genes And Genomes Flashcards
Recombinant DNA
Depends on having enzymes that cut, join and replicate. DNA ligament join them together
Recombinant DNA is used to
Construct new combinations of DNA as well as to detect and amplify particular sequences
Southern and northern blots
Used to separate and amplify DNA and RNA sequences
Restriction enzymes
Also called restriction endonucleases
Recognize specific base sequences in double helical DNA
Palindromic sequence
Cleave sites that are symmetrically positioned
Break phosphodiester bonds
Southern blotting
Specific sequence separated by electrophoresis, transferring to nitrocellulose and hydrolyzing with P labeled probe
Sanger dideoxy method
Controlled termination of replication
ddNTP blocks termination because it has an H instead of an OH
DNA oligonucleotide synthesis
Works backwards
Contains DMT which protects 5’O atom and has BCE protecting the 3’
NH3 is added to remove all protecting groups
PCR
Need a pair of primers All dNTPS A heat stable DNA polymerase Done in vitro 2^n fold
Vitro method for DNA replication
Making DNA cloning Screwing DNA to check its size Fingerprinting Mutagenesis Real time PCR
Plasmid vecto
DNA molecule that can replicate autonomously in an appropriate host organism
DNA ligament
Requires ATP
Catalyzes covalent bond
Creates phosphodiester bond
cDNA
Formed by reverse transcriptase
Sticks to oligo(T) primer
We can mutate something if we know
The base sequence around the site to be altered
Have a plasmid containing the cDNA for the protein
Study gene transcription
Individual mRNA can be quantified by qPCR
cDNA is prepared
How quick did each sample take to get to the threshold?
