Lab 4 – DNA purification and RFLP

Question 1

Many enzymatic reactions fail to work properly

Answer 1

the enzyme is exposed to the wrong combination of ions or is exposed to an excess of contaminating protein.

Question 2

can interfere with other enzymes that require different buffering systems

Answer 2

The presence of one type of enzyme/buffer (e.g. Taq polymerase)

Question 3

are prokaryotic-derived enzymes commonly used to “chop-up” larger DNA molecules into smaller fragments.

Answer 3

Restriction endonuclease enzymes

(also known as “restriction enzymes”)

Question 3

Bacteria naturally use these enzymes as a way of destroying foreign DNA and protecting themselves from viral intruders

Answer 3

Restriction endonuclease enzymes

(also known as “restriction enzymes”)

Question 4

cut DNA in a very sequence-specific manner (rather than blindly digesting any DNA).

Answer 4

Restriction enzymes

Question 5

only cuts DNA when it sees the sequence 5’-GAATTC-3’

Answer 5

restriction enzyme EcoRI

Question 6

Most restriction enzyme recognition sites resemble that of EcoRI in that most are only what nucleotides long

Answer 6

4-6

Question 7

are those that have the same sequence on both strands if read 5’ –> 3’.

Answer 7

palindromic

Question 8

When you read both strands 5’3’ you get the same GAATTC sequence

Answer 8

palindrome

Question 9

any two pieces of DNA cut with the same restriction enzyme can be

Answer 9

artificially glued together

Question 10

it is possible to combine pieces of DNA that are derived from completely different sources into a brand new type of DNA

Answer 10

recombinant DNA

Question 11

single nucleotide differences

Answer 11

polymorphisms

Question 12

This technique of analyzing differences in DNA sequence using restriction enzymes

Answer 12

RFLP analysis

Question 13

Before DNA sequencing was readily available, the preferred way of analyzing DNA differences was via

Answer 13

RFLP

Question 14

serve as a vital tool for detecting sequence differences in the genomes of individuals within a population

Answer 14

restriction enzymes

Question 15

contains chaotropic salts (e.g. guanidinium chloride, urea) that function to denature all proteins agent. In other words, these are chemical denaturing agents.

Answer 15

Buffer PB

Question 16

contains salts and a high concentration of ethanol (70%). Just like the first lab, 70% ethanol is very commonly used as a DNA washing agent

Answer 16

Buffer PE