Tools and techniques

Question 1

Basic requirements of nucleic acids isolation

Answer 1

Disruption, separation/isolation/deproteinisation, purification

Question 2

Detergent for plant cells

Answer 2

CTAB

Question 3

Detergent for animal, yeast, bacteria and fungi cells

Answer 3

SDS

Question 4

Reactants used for deproteinisation of plan cells

Answer 4

Chloroform/isoamyl alcohol

Question 5

Reactants used for deproteinisation of animal and bacterial cells

Answer 5

Phenol/chloroform

Question 6

Once deproteinisation occurs, where’s located nucleic acids?

Answer 6

In the supernant, acquous part.

Question 7

If you want to get only mRNA what should you add?

Answer 7

oligo dT-cellulose

Question 8

When precipitating genetic material, what do you add to the material?

Answer 8

Cold isopropanol (or ethanol), with high concentration of salt, for protecting DNA. Then washed with 70% Ethanol and washed with TE (Tris EDTA) to takes out the metal of the salt. And fially to get rid of RNA, and RNase.

Question 9

What’s a chelant molecule, example and how it helps you when precipitating nucleic acids?

Answer 9

This molecule takes metals, example TE, and helps you to take out metals from salts previously added.

Question 10

Method used for mRNA purification

Answer 10

Affinity chromatography

Question 11

Method used for plasmid DNA purification

Answer 11

Gradient centifugation

Question 12

Chemical used for pDNA purification

Answer 12

CsCl

Question 13

For long storage of DNA, what buffer is recommended?

Answer 13

TE (Tris-EDTA)

Question 14

Absorbance of of dsDNA at 260nm

Answer 14

50microg/mL

Question 15

Absorbance of of ssDNA/RNA at 260nm

Answer 15

40microg/mL

Question 16

Ratio of wavelength for determining purity

Answer 16

260/280 nm between 1.8-2

Question 17

Isotopes used for dNTPs at radiolabelling

Answer 17

3H, 32P, 35S, 14C

Question 18

Differece between probing and tracking

Answer 18

Specificity

Question 19

Radiolabelling that uses Polynucleotide kinase

Answer 19

End labelling

Question 20

Purines

Answer 20

A y G

Question 21

Pirimidines

Answer 21

T y C

Question 22

What does PNK does?

Answer 22

In end labelling, puts a radioactive P onto the 5’ hydroxi termini

Question 23

What enzyme is used in nick translation

Answer 23

DNA Pol. I

Question 24

How do you create a nick in the DNA

Answer 24

Low concentration DNase

Question 25

What’s first done when prime labelling?

Answer 25

Denature DNA by heating

Question 26

What does it neans that DNA is plyanionic?

Answer 26

It has many negative charges

Question 27

Method used for protein electrophoresis

Answer 27

SDS-PAGE

Question 28

Chemical used for gelification of plyacrilamide

Answer 28

TEMED

Question 29

Buffer used for SDS-PAGE

Answer 29

TG

Question 30

What beta mercaptoethanol does?

Answer 30

Brakes S-S bonds in proteins

Question 31

Name of the dye in electrophoresis

Answer 31

Bromophenol blue

Question 32

Standard cencentration of agarose and voltage in SAGE

Answer 32

1%, 120V