Lab Flashcards
What are the 3 steps of PCR?
1) denaturation (hot)
2) annealing (cool)
3) extension/synthesis (hot)
What does the denaturation step of PCR entail?
splits DNA strands
What does the annealing step of PCR entail?
primers bind to DNA strands
What does the extension/synthesis step of PCR entail?
primers elongated
What is the function of the Taq polymerase in PCR? Why is a bacteria like E coli not used?
elongates primers added during annealing by adding complementary nucleotides; thermophilicity of exremophile required
PCR Reaction Component: Taq polymerase
Function: ?
adds complementary nucelotides
PCR Reaction Component: PCR buffer
Function: ?
stabilize DNA and other components
PCR Reaction Component: Primer
Function: ?
short DNA sequences that allow Taq to elongate DNA
PCR Reaction Component: dNTPs
Function: ?
complementary nucleotides added by Taq used to elongate primers
PCR Reaction Component: MgCl2
Function: ?
required for Taq polymerase
PCR Reaction Component: ? Function: adds complementary nucelotides
Taq polymerase
PCR Reaction Component: ?
Function: stabilize DNA and other components
PCR buffer
PCR Reaction Component: ?
Function: short DNA sequences that allow Taq to elongate DNA
Primer
PCR Reaction Component: ?
Function: required for Taq polymerase
MgCl2
Formula for total copies created using PCR
DNA(total)=DNA(copies)x2^n
n=# of cycles
How are DNA fragments separated using agarose gel electrophoresis?
DNA fragments are separated based on charge and size (small move quicker)
What is the purpose of adding ethidium bromide to the gel?
binds to DNA and flueorscences udner UV light
Why was sample loading buffer added to your samples before loading it into the well in the agarose gel?
To increase the desnity of DNA fragments to prevent mizing DNA with the buffer
How can you determine the molecular weight of DNA fragments using agarose gel electrophoresis?
smaller fragments move faster than larger fragments
What is taxonomy?
the science of namin and grouping organisms based on genetic relationships?
How does taxonomy differ for eukaryotes vs prokaryotes?
eukaryotes- sexual reproduction
prokaryotes- similar characteristic
The identification of bacteria is aided by a multivolume identification manual known as
Manual of Systematic Bacteriology
What is the Manual of Systematic Bacteriology?
multi-volume manual use in the identification of bacteria
Identification of bacteria typically begins with ____
gram stain and morphological characteristics
A catalase test can be used to separate ____ bacteria from ____ bacteria
gram positive; gram negative
