Extraction purification

Question 1

the 1st step in DNA isolation

Answer 1

Lysis

Question 2

Anticoagulant used in Ficoll directed Gradient

Answer 2

Ethylenediaminetetraacetic acid or citrate

Question 3

solation of peripheral blood mononuclear cells and plasma are obtained by centrifugation and addition by density gradient medium called

Answer 3

Ficoll hypaque

Question 4

Whole blood mixed with Tris-HCL (pH 8.0), saturated phenol, and water
followed by constant shaking for 4 hours at room temperature.

Answer 4

phenol

Question 5

Whole blood mixed with Tris, EDTA, sodium dodecyl sulfate, MgCl2, and
proteinase K in presence of high salt for overnight digestion at 37ºC.

Answer 5

proteinase K

Question 6

is the most commonly used lysing agent

Answer 6

Detergent

Question 7

also known as zymolase, can be used to lyse fungal cell
walls.

Answer 7

Lyticase

Question 8

Techniques used for purification DNA

Answer 8

Organic
Inorganic
Spin column

Question 9

Four Steps in solid phase includes the ff

Answer 9

1. Cell lysis
2. Nucleic acid absorption
3. Washing
4. Elution

Question 10

Mixture of Phenol: Chloroform: Isoamyl alcohol ratio used
to inhibit activity of RNase activity

Answer 10

25:24:1

Question 11

after centrifugation what layer contains DNA in organic method

Answer 11

Upper

Question 12

Removal of unbind particles or proteins

Answer 12

Washing

Question 13

step that uses RNase free water is added for release of nucleic acid from silica membrane

Answer 13

Elution