Biomed PCR & Lab quiz

What does PCR stand for

Polymerase chain reaction

PCR Goal

Amplify (make copies) a specific target DNA sequence

PCR Machine Name

Thermal Cycler

Enzyme added to PCR reaction and function

Taq polymerase, adds nucleotides to extend DNA

Building blocks to make more DNA

Nucleotides (dNTPs)

buffer purpose

to keep optimal pH

What helps tell polymerase where to start copying and how many do we need?

primer, 2 (forward & reverse)

PCR time length

2-4 hours

positive control purpose

provide a visual example, ensure the PCR is working

Why is taq polymerase special and where discovered?

high temp resistance and discovered in Yellowstone geyser bacteria

Steps of PCR

Denaturation (95 degrees & H bonds breaking between base pairs) → Annealing (55 degrees & primer attachment) → Extension (75 degrees & strand extension with taq)

tt

non-taster Gene (1 band 221 bp)

Tt

Slight taster (3 bands 221bp 176bp & 45 bp)

TT

Super taster gene (2 bands 176bp & 45 bp)

SNP

Single Nucleotide Polymorphism (base change that can cause a change in amino acid)

how many SNPs in TAS2R38 gene

3

What should you do if you have a small amount of DNA and want to make copies of it?

PCR

What restriction enzyme did the lab use and what was its function?

Hae III, blunt cut DNA into fragments that are each different sizes

Hae III Restriction site

GGCC

what does Hae III cut

the taster variant (TT & Tt)

which fragments move farthest in gel electrophoresis

the shortest

what liquid had the DNA

the supernatant

DNA charge

Negative

what side of the Gel should the DNA be loaded in

the negative side