Extraction and Purification of nucleic acids-Proteins

Question 1

Is the total set of proteins encoded by genome

Answer 1

Proteome

Question 2

Proteins can be extracted from
various sources, including:

Answer 2

Microorganism
Animal cell or tissue

Question 3

Separates soluble proteins from cell debris

Answer 3

Centrifiguration

Question 4

Removes particulates and aggregates

Answer 4

Filtration

Question 5

Exploits solubility differences

Answer 5

Salting out (Ammonium sulfate precipitation)

Question 6

Uses pH to precipitate proteins at their isoelectric point.

Answer 6

Isoelectric precipitation

Question 7

Separates proteins based on their size

Answer 7

Size exclusion chromatography

Question 8

Separates proteins based on charge

Answer 8

Ion exchange chromatography

Question 9

Uses a ligand that specifically binds to the target protein.

Answer 9

Affinity Chromatography

Question 10

Exploits hydrophobicity differences

Answer 10

Hydrophobic interaction chromatography

Question 11

Protocols are generally employed when the sample of interest consist of cells that lyse easily, such as red blood cells and tissue culture cells

Answer 11

Gentle cell distruption

Question 12

Cell swell and burst when they are suspended in hypotonic solution, releasing all cellular components.

Answer 12

Osmotic lysis

Question 13

cells can be lysed by subjecting them to one or more cycles of quick freezing using liquid nitrogen and subsequent thawing

Answer 13

Freeze thaw lysis

Question 14

cellular membranes can be
solubilized just by suspending the cells in many detergent-containing solutions.

Answer 14

Detergent lysis

Question 15

by enzymes that specifically digest the cell wall.

Answer 15

,Isosmotic solution

Question 16

What are the enzyme present in the isosmotic solution

Answer 16

Cellulase
Pectinase for plant cells
Lyticase for yeast cells
Lysozyme for bacterial cells

Question 17

Biological material with tough cell walls and many tissue types require more rigorous methods, which are mainly based on mechanical rupture.

Answer 17

Sonication
French press
mortar and pestle
Mechanical homogenization

Question 18

A cell suspension, cooled on ice to avoid heating, can be distrupted by shear forces using short burst of ultrasonic waves

Answer 18

Sonication

Question 19

Cells are lysed by shear force by forcing a cell suspension through a small orifice at high pressure

Answer 19

French press

Question 20

Breaking down cells of solid tissues and microorganisms.

Answer 20

Mortar and pestle

Question 21

Soft, solid, tissues can be employed using handheld devices, blenders, or other motorized devices

Answer 21

Mechanical Homogenization

Question 22

Can also be employed for cell distruption.

Answer 22

Glass bead homogenization

Question 23

The uses of precipitation vary based on three (3) main
factors:

Answer 23

Detergent of contanimants for removal
Native or denatured state of proteins
Post processing analysis

Question 24

This precipitation method is a
concentration-based procedure.

Answer 24

Acetone precipitation

Question 25

Protein solution is mixed with a dilute sodium deoxycholate buffer

Answer 25

TCA (Trichloroacetic acid) Precipitation

Question 26

precipitation is remarkably efficient, even in dilute samples, with recovery values from 90% to 100% of total protein.

Answer 26

TCA-DOC

Question 27

method of precipitation and concentration for dilute samples, especially those containing membrane proteins.

Answer 27

Methanol chloroform precipitation

Question 28

The ratio of methanol:chloroform: water is

Answer 28

4:1:3

Question 29

Analyzes protein purity and molecular weight

Answer 29

SDS PAGE (Sodium dodecyl sulfate polyacrylamide gel electrophoresis)

Question 30

Identifies proteins using antibodies

Answer 30

Western blot

Question 31

Determines protein mass and structure

Answer 31

Mass spectrometry

Question 32

measures protein concentration

Answer 32

UV-VIS Spectroscopy (280 Nm)

Question 33

Assesses functionality if the protein is an enzyme

Answer 33

Enzyme activity assays