PCR

PCR

molecular technique used to amplfies DNA by using specfic primers & TAQ

GMO

genitically modified organism

35s promoter

drives the expression of GMO genes

What are the componets inside PCR?

Taq polymerase, forward/reverse primers,nucelotide, dna, DNTPS, buffers

GMO+

35s promoter

GMO-

no 35s promoter

tublin

Protein for eukaryotes & to see if DNA extraction & amplfication worked correctly

gel star

agent that binds to dDNA & enables visulzations under UV light

what is gel electrophoresis ?

seperates molecules by size

what are the steps of PCR?

denaturation, elongation, and annealing

1x TBE buffer

buffer used for gel electrophoresis

Whats the purpose of molecular ladder?

known vs unkoiwn sizes of DNA fragments

denaturation ? temperture

95 ; seperates double helix into single stradn to use for template

Annealing ? temp?

55-65, allows primers to bind to sepcfic sequances in target DNA template

elongation? temp?

72 ; allows TAQ polyermase, enzyme to build a new DNA strand

Whats the prupose of TAq polymerase?

enzyme that makes copies of specfic DNA sequances during PCR *resistant to high tempertures

What are the 4 stages of DNA extraction?

1)cell lysis 2) DNA precipation 3) DNa purification 4)DNA resuspension

isporpropyl?

preicpates DNA

ethanol?

pueifes dna

edwards buffer

lysing buffer

tTRNA

carries out gel elctroppherosis and resusspenison

DNA is replicated from the 5’-3’ end and TAQ polymerase is added from the 3’ end. T/F?

trueee

should the wells be closest to the black electrodes?

yes

gel eelctrophoresis seperates based on?

size

Tubulin is an internal control and therefore
a band should be detected in ALL samples, true/false

trueeee

what are types of DNTPS?

dGTP, dCTP, dTTP, dATP,