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Gel electrophorisis
separates DNA fragments by size, using electric currents, to compare to other DNA
Polymerase Chain reaction
replicates DNA in 3 steps. Denaturation, Annealing and Elongation
Denaturation
part of PCR heat DNA to 94 degrees so the hydrogen bonds brake separating the DNA into 2 strands
annealing
Part of PCR reduces temperature to 55 degrees so the primers can attach
Elongation
part of PCR temperature increase to 72 degrees so the Taq polimerase works at it ideal temp so it most efficient
what you need for PCR
• DNA • Primers • Necleotides • Taq polimerase • mix buffer • PCR tube
What charge is DNA
Negative (-)
what charge does DNA move to
positive terminal
what “glues” the DNA backbone together after it has been cut
Ligases
whats responsible for “cutting” DNA
restriction enzymes
standard ladder
use in gel electrophoresis to compare to the unknown sample
polimerse
add complimentry nucleotides to a sequence.
CRISPR cas-9 in Bacteria
Cause viral DNA to become non functioning
CRISPR Cas-9 in genetic engineering
causes a chosen gene thats already been created to go in the cut site
cleaves
CRISPER Cas-9 cuts DNA
Exposure
bacteria is exposed to viral DNA it takes a sample of the DNA and put it in the PAM sequence.
Expression
cas-9 and guide RNA (from PAM sequence) combine making a CRISPR cas-9 complex, which scan the DNA for a complimentary to gRNA
Extermination
If complimentry bases to guide RNA are found the cas-9 cleaves (cuts) it and random necleotides make the viral DNA non-functional
what type or RNA is in a CRISPR Cas-9 complex?
guide RNA, gRNA