Biotechnological Tools Flashcards
Where are restriction enzymes cut
Restriction sites
What are blunt ends and sticky ends
Blunt - evenly broken
Sticky - unevenly broken
How are they re-annealed
DNA ligase joins them
(T4 works on blunt ends)
How is it useful (RESTRICTION)
They modify the DNA to make it more useful by splicing and inserting segments of DNA
Outline gel electrophoresis
It separates large molecules like DNA, RNA and proteins
Prepare gel and place between electrodes
Load DNA sample solutions into wells of gel
Apply an electric current
Stain the cell to see DNA fragments
Why does DNA travel toward positive electrode
They are negatively charged fragments
Which travel furthest, Why?
the smaller fragments so that the DNA fragments can be separated
How is it useful (GEL)
You can distinguish DNA fragments of different lengths.
How is human gene spliced
Restriction enzymes
How is plasmid inserted into bacterial cell
A host cell
How is it efficient (Insulin)
What does PCR stand for
Polymerase Chain Reaction
PCR purpose
makes a huge number of copies of a DNA sequence quickly and without a host organism
How is PCR different from DNA replication
PCR does targeted amplification to replication only a segment of DNA bounded by the two primers. DNA amplifies all of the cell’s DNA/
Plasmid mapping
graphical representation of plasmids, that show the locations of major identifiable landmarks on DNA like restriction enzyme sites, gene of interest, plasmid name and length etc.
