Basic Methods And Tools In Molecular Biology And Diagnostics

Question 1

This is the most critical part in molecular techniques because downstream procedures and analysis relies on the quality of the DNA used for the assay.

Answer 1

DNA extraction

Question 2

This refers to the process of separating DNA from other cellular materials such as CHONS and membranes

Answer 2

DNA extraction

Question 3

DNA extraction refers to the process of separating DNA from other cellular materials such as _____ and _____.

Answer 3

CHONS
membranes

Question 4

What do DNA extraction remove?

Answer 4

Potential inhibitors

Question 5

After the removal of potential inhibitors to the PCR amplification, what does the DNA extraction produce?

Answer 5

Stable solution of high quality DNA

Question 6

What are the 3 basic steps of specimen preparation DNA and RNA extraction?

Answer 6

Lysis
Precipitation
Purification

Question 7

This step of specimen preparation DNA and RNA extraction has the cell and nucleus broken open to release the DNA inside.

Answer 7

Lysis

Question 8

In the lysis step, what is broken open to release the DNA inside?

Answer 8

Cell and nucleus

Question 9

Under the lysis step of specimen preparation DNA and RNA extraction, what are the methods used?

Answer 9

Mechanical disruption and using the detergents and enzymes such as:
Organic: NaOH, SDS
Inorganic: Tris-EDTA (TE), EDTA, SDS

Question 10

This step of specimen preparation DNA and RNA extraction, this separated DNA from cellular debris

Answer 10

Precipitation

Question 11

Under the precipitation step of specimen preparation DNA and RNA extraction, the DNA is separated from what?

Answer 11

Cellular debris

Question 12

This neutralizes the negative charges on the DNA molecules which makes them more stable and less water soluble.

Answer 12

Sodium ions

Question 13

Sodium neutralizes the negative charges on the DNA molecules which makes them _____ and _______.

Answer 13

more stable
less water soluble.

Question 14

Under the precipitation step, this is added which causes DNA to precipitate out the aqueous solution because it is not soluble in alcohol.

Answer 14

Ethanol/isopropanol

Question 15

What is not soluble in ethanol?

Answer 15

DNA

Question 16

Under purification, the extracted DNA from precipitation can be rinsed with _______ to remove any remaining unwanted material and cellular debris.

Answer 16

Alcohol

Question 17

What are the examples of cell lysis buffer?

Answer 17

Non-ionic detergent, salt, buffer and EDTA

Question 18

This is designed to lyse outer cell-membrane but will not breakdown the nuclear membrane.

Answer 18

Cell lysis buffer

Question 19

The cell lysis bufferis designed to lyse _______ but will not breakdown the ________?

Answer 19

outer cell-membrane
nuclear membrane

Question 20

When this reagent for extraction is used the nucleus is intact.

Answer 20

Cell lysis buffer

Question 21

This is used to remove contaminating protein

Answer 21

Phenol

Question 22

This prevents cutting of DNA during isolation

Answer 22

Chloroform

Question 23

What does the chloroform prevent?

Answer 23

Cutting of DNA during isolation

Question 24

This reagent for extraction solubilizes lipids and a lot of protein to remove them from the DNA.

Answer 24

Chloroform

Question 25

Chloroform solubilizes ______ and a lot of _______ to remove them from the DNA.

Answer 25

lipids protein

Question 26

This removes most of the CHON by digesting with proteolytic enzyme

Answer 26

Proteinase K

Question 27

This is against a broad spectrum of native proteins and denatured proteins

Answer 27

Proteinase K

Question 28

This is a chelating agent of magnesium and cofactor of DNase

Answer 28

EDTA

Question 29

If the magnesium is bound to EDTA, DNase is inactivated. Therefore, protecting _____ from damage

Answer 29

DNA

Question 30

This is an anionic detergent which helps cell membrane and nuclear envelope to break open

Answer 30

Sodium Dodecyl Sulfate (SDS)

Question 31

This inhibits RNAse and DNAse

Answer 31

Sodium dodoceyl sulfate (SDS)

Question 32

What kind of reagent is SDS?

Answer 32

Anionic detergent

Question 33

What does the SDS help to break open?

Answer 33

Cell membrane Nuclear envelope

Question 34

This prevents the DNA and RNA degradation

Answer 34

Tris-EDTA buffer

Question 35

This is used for final resuspension and storage because it preserves any change in pH

Answer 35

Tris-EDTA buffer

Question 36

This reagent is used for the precipitation of DNA

Answer 36

95% ethanol or isopropanol

Question 37

What are the different types of DNA extraction methods?

Answer 37

Chemical based DNA extraction method Solid phase DNA extraction method

Question 38

What are the types of CBDNAEM?

Answer 38

Organic and inorganic solvent based DNA extraction extraction

Question 39

What is used under organic-base DEM?

Answer 39

Phenol chloroform method

Question 40

What is used under inorganic-based DEM?

Answer 40

SALTING OUT METHOD PROTEINASE K METHID/ ENZYMATIC METHOD