BIOL234Z Methods in biology

Question 1

What does RFLP stand for?

Answer 1

Restriction fragment length polymorphism

Question 2

What is the function of a restriction enzyme?

Answer 2

A protein isolated from bacteria that cleaves DNA sequences at specfic sites

Question 3

What methods can be used to identify MT-CYB?

Answer 3

Agarose gel, spectophotometry

Question 4

What is the central dogma?

Answer 4

theory that genetic information only flows in one direction

Question 5

During specto what level does DNA absorb at?

Answer 5

260

Question 6

Why do we have to extract DNA?

Answer 6

1. break down cell walls/membranes
2. remove all cellular material except DNA
3. clean and resuspend DNA either in water or a low conc. buffer suitable for storage

Question 7

What are the steps needed to complete a PCR?

Answer 7

Template (DNA)
Forward and reverse primers
dNTPS
DNA polymerase
Buffer & catalyst

Question 8

What are the stages of PCR?

Answer 8

Denaturation
Annealing
Extension/Polymerase

Question 9

What is the c- vaue paradox?

Answer 9

The amount of DNA not corresponding with the complexity of an organism

Question 10

What is the function of restriction enzymes?

Answer 10

molecular scissors

Question 11

What is the sanger method?

Answer 11

Selective incorporation of dideonucleotides (ddNTPs) that terminate chain elongation

Question 12

What are microsatelites?

Answer 12

Small pieces of DNA that only are a few base pair repeating lengths

Question 13

From which pole would DNA run from in agarose gel?

Answer 13

Negative to positive

Question 14

What is AFLP and what is it used for?

Answer 14

amplified length polymorphism, amplification of a section of DNA fragments to generate and compare.

Question 15

What is SNPs and what is it used for?

Answer 15

Single Nucleotide Polymorphisms; used to detect a genomic variant at a single base position in the DNA

Question 16

Name 2 techniques that are considered 3rd generation sequencing

Answer 16

Oxford Nanopore sequencing and PacBio SMRT sequencing

Question 17

What are the reagents needed to complete Sanger sequencing?

Answer 17

Template DNA
Forward primer
dNTPs
DNA polymerase
Buffers and catalysts
ddNTPs

Question 18

State the general steps in Sanger sequencing

Answer 18

1. Prepare the reaction mixture which includes: Primer and DNA template, DNA polymerase, and ddNTPs with fluorochromes.
2. Primer elongation and chain termination
3. Capillary gel electrophoresis separation of DNA fragments
4. Laser detection of fluorochromes and computational analysis of sequence.