Unit 1: Genomic DNA from plant - Practice Quiz

BTY555 — Biotechnology Laboratory-I 60 Questions
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1 What does the abbreviation CTAB stand for?

Extraction of genomic DNA from plant by using cTAB method Easy
A. Cetyl Trimethyl Ammonium Bromide
B. Chloroform Tris Ammonium Buffer
C. Cetyl Tri Amino Butyrate
D. Calcium Tetra Ammonium Bromide

2 The CTAB method is primarily used to extract which type of biomolecule?

Extraction of genomic DNA from plant by using cTAB method Easy
A. Genomic DNA
B. Membrane lipids
C. Ribosomal RNA
D. Storage proteins

3 What is the chemical nature of CTAB detergent?

Extraction of genomic DNA from plant by using cTAB method Easy
A. Non-ionic detergent
B. Cationic detergent
C. Anionic detergent
D. Zwitterionic detergent

4 Why is the CTAB method especially preferred for plant tissues?

Extraction of genomic DNA from plant by using cTAB method Easy
A. It sequences the extracted DNA
B. It amplifies target DNA sequences
C. It stains the DNA for visualization
D. It removes polysaccharides and polyphenols effectively

5 Which technique is commonly used to break down the rigid plant cell wall before adding CTAB buffer?

Extraction of genomic DNA from plant by using cTAB method Easy
A. Grinding tissue in liquid nitrogen
B. Boiling in distilled water
C. Sonication in ethanol
D. Freezing at

6 What is the typical role of -mercaptoethanol in the CTAB extraction buffer?

Extraction of genomic DNA from plant by using cTAB method Easy
A. Cuts the DNA into fragments
B. Digests contaminating proteins
C. Precipitates the DNA
D. Reduces oxidation of polyphenols

7 What is the main function of EDTA in the CTAB extraction buffer?

Extraction of genomic DNA from plant by using cTAB method Easy
A. Denatures the DNA strands
B. Neutralizes the buffer pH
C. Chelates metal ions and inhibits DNase
D. Precipitates the polysaccharides

8 The organic solvent mixture chloroform:isoamyl alcohol is used mainly to:

Extraction of genomic DNA from plant by using cTAB method Easy
A. Lyse the cell membrane
B. Separate proteins from nucleic acids
C. Buffer the solution pH
D. Precipitate the genomic DNA

9 In the chloroform:isoamyl alcohol mixture, what is the usual ratio used?

Extraction of genomic DNA from plant by using cTAB method Easy
A. 24:1
B. 3:2
C. 10:1
D. 1:1

10 After centrifugation of the aqueous phase, which reagent is commonly added to precipitate the DNA?

Extraction of genomic DNA from plant by using cTAB method Easy
A. Acetic acid
B. Sodium hydroxide
C. Distilled water
D. Chilled isopropanol

11 During DNA precipitation, why is the sample often kept at low temperature?

Extraction of genomic DNA from plant by using cTAB method Easy
A. To enhance DNA precipitation
B. To dissolve polysaccharides
C. To break the DNA strands
D. To activate DNases

12 Which alcohol concentration is generally used to wash the DNA pellet to remove salts?

Extraction of genomic DNA from plant by using cTAB method Easy
A. 70% ethanol
B. 10% ethanol
C. 50% isopropanol
D. 100% methanol

13 What is the purpose of incubating the ground tissue with CTAB buffer at around ?

Extraction of genomic DNA from plant by using cTAB method Easy
A. To amplify DNA fragments
B. To digest the DNA
C. To sequence the genomic DNA
D. To promote cell lysis and DNA release

14 Which enzyme is often added to the extracted DNA to remove RNA contamination?

Extraction of genomic DNA from plant by using cTAB method Easy
A. Proteinase K
B. Ligase
C. DNase
D. RNase

15 In the CTAB protocol, what is commonly used to finally dissolve and store the purified DNA?

Extraction of genomic DNA from plant by using cTAB method Easy
A. CTAB buffer
B. Chloroform
C. TE buffer
D. Concentrated ethanol

16 Which technique is commonly used to check the quality and integrity of extracted genomic DNA?

Extraction of genomic DNA from plant by using cTAB method Easy
A. Mass spectrometry
B. Titration
C. Agarose gel electrophoresis
D. Flame photometry

17 The purity of extracted DNA is commonly assessed using the absorbance ratio at:

Extraction of genomic DNA from plant by using cTAB method Easy
A.
B.
C.
D.

18 A high concentration of NaCl in the CTAB buffer mainly helps to:

Extraction of genomic DNA from plant by using cTAB method Easy
A. Keep polysaccharides soluble and separate from DNA
B. Neutralize -mercaptoethanol
C. Break the plant cell wall
D. Degrade the genomic DNA

19 At what wavelength does DNA show maximum absorbance, used for quantification?

Extraction of genomic DNA from plant by using cTAB method Easy
A. 320 nm
B. 280 nm
C. 260 nm
D. 230 nm

20 During phase separation, in which layer is the genomic DNA found?

Extraction of genomic DNA from plant by using cTAB method Easy
A. Lower organic phase
B. Surface foam layer
C. Interphase debris
D. Upper aqueous phase

21 In the CTAB method of plant DNA extraction, what is the primary function of the detergent CTAB (cetyltrimethylammonium bromide)?

Extraction of genomic DNA from plant by using cTAB method Medium
A. To solubilize membrane lipids and form complexes with polysaccharides
B. To digest all cellular RNA molecules
C. To precipitate DNA directly from the cell
D. To denature the double-stranded DNA into single strands

22 Why is a high salt concentration (typically NaCl) maintained in the CTAB extraction buffer?

Extraction of genomic DNA from plant by using cTAB method Medium
A. It precipitates proteins immediately
B. It activates nucleases to remove RNA
C. It keeps CTAB-polysaccharide complexes soluble while allowing DNA to remain in solution
D. It lowers the pH to protect DNA

23 What is the role of -mercaptoethanol when added to the CTAB extraction buffer?

Extraction of genomic DNA from plant by using cTAB method Medium
A. It degrades contaminating RNA
B. It reduces disulfide bonds and inhibits oxidation of phenolic compounds
C. It increases the ionic strength of the buffer
D. It precipitates the genomic DNA

24 During CTAB extraction, a chloroform:isoamyl alcohol (24:1) mixture is used. What is the purpose of the isoamyl alcohol component?

Extraction of genomic DNA from plant by using cTAB method Medium
A. It precipitates the DNA into the organic phase
B. It increases the density of the aqueous phase
C. It digests residual proteins enzymatically
D. It reduces foaming and stabilizes the interphase between aqueous and organic layers

25 Why is plant tissue typically ground in liquid nitrogen before adding CTAB buffer?

Extraction of genomic DNA from plant by using cTAB method Medium
A. To break rigid cell walls into a fine powder while minimizing nuclease and enzyme activity
B. To sterilize the sample completely
C. To dissolve the polysaccharides directly
D. To precipitate the DNA before lysis

26 After the chloroform extraction step, in which phase is the genomic DNA found?

Extraction of genomic DNA from plant by using cTAB method Medium
A. The upper aqueous phase
B. The white interphase
C. The lower organic phase
D. Distributed equally in both phases

27 Isopropanol (or chilled ethanol) is added to the recovered aqueous phase in the CTAB protocol. What does this step accomplish?

Extraction of genomic DNA from plant by using cTAB method Medium
A. It precipitates the DNA out of solution
B. It degrades contaminating RNA
C. It removes CTAB by binding to it
D. It solubilizes remaining polysaccharides

28 Why is a ethanol wash performed on the DNA pellet after precipitation?

Extraction of genomic DNA from plant by using cTAB method Medium
A. To digest any co-precipitated proteins
B. To dissolve the DNA completely for measurement
C. To remove residual salts and CTAB while keeping the DNA precipitated
D. To denature the double-stranded DNA

29 A student's extracted DNA has an ratio of . What does this most likely indicate?

Extraction of genomic DNA from plant by using cTAB method Medium
A. Pure, high-quality DNA
B. Protein or phenolic contamination in the sample
C. RNA contamination only
D. Excessive salt in the sample

30 Why is the CTAB method particularly preferred over standard SDS-based methods for many plant tissues?

Extraction of genomic DNA from plant by using cTAB method Medium
A. It requires no organic solvents at all
B. It works only at low salt concentrations
C. It avoids the need for grinding tissue
D. It effectively removes polysaccharides that co-purify with plant DNA

31 Polyvinylpyrrolidone (PVP) is often added to the CTAB buffer for plants rich in phenolics. What is its function?

Extraction of genomic DNA from plant by using cTAB method Medium
A. It degrades contaminating RNA
B. It binds and removes polyphenolic compounds
C. It precipitates the genomic DNA
D. It raises the salt concentration of the buffer

32 The CTAB extraction buffer is commonly incubated at –. What is the main reason for this incubation?

Extraction of genomic DNA from plant by using cTAB method Medium
A. To denature the DNA into single strands
B. To precipitate the DNA at high temperature
C. To enhance cell lysis and inactivate DNases while promoting CTAB action
D. To evaporate residual chloroform from the mix

33 EDTA is a common component of CTAB extraction buffers. Why is it included?

Extraction of genomic DNA from plant by using cTAB method Medium
A. It removes polysaccharides from solution
B. It chelates ions, inhibiting DNase activity
C. It precipitates the DNA rapidly
D. It reduces oxidation of phenolic compounds

34 Tris-HCl (usually at pH ) is a component of the CTAB buffer. What role does it play?

Extraction of genomic DNA from plant by using cTAB method Medium
A. It maintains a stable, slightly alkaline pH that protects DNA from degradation
B. It acts as the primary reducing agent
C. It precipitates proteins from the lysate
D. It supplies the high salt needed to remove polysaccharides

35 To obtain RNA-free genomic DNA using the CTAB method, what treatment is typically applied?

Extraction of genomic DNA from plant by using cTAB method Medium
A. Additional CTAB precipitation
B. Treatment with RNase A
C. A second chloroform extraction step to physically pull the RNase A enzyme through the aqueous interphase while simultaneously removing all residual polysaccharides and salts from the sample
D. An extra ethanol wash

36 A student sees a bright band of intact high molecular weight DNA but also a strong smear at the bottom of an agarose gel. What does the smear most likely indicate?

Extraction of genomic DNA from plant by using cTAB method Medium
A. Complete absence of DNA
B. Presence of high molecular weight DNA only
C. Pure, undegraded genomic DNA
D. Degradation of the DNA into smaller fragments

37 During DNA precipitation, the sample is often incubated at . What is the benefit of this step?

Extraction of genomic DNA from plant by using cTAB method Medium
A. It denatures the DNA into single strands
B. It removes phenolic contaminants
C. It degrades residual RNA
D. It improves the yield by enhancing DNA precipitation

38 Why should vigorous vortexing be avoided during the CTAB extraction of genomic DNA?

Extraction of genomic DNA from plant by using cTAB method Medium
A. It denatures the RNase enzyme
B. It lowers the salt concentration in the buffer
C. It causes mechanical shearing of the high molecular weight DNA
D. It prevents CTAB from binding polysaccharides

39 The purified DNA pellet is finally dissolved in TE buffer rather than distilled water. Why is TE buffer preferred for storage?

Extraction of genomic DNA from plant by using cTAB method Medium
A. TE degrades any contaminating RNA in the sample
B. TE lowers the pH to acidic levels for stability
C. The EDTA in TE chelates divalent cations and inhibits any residual DNase activity during long-term storage
D. TE precipitates the DNA to keep it stable

40 If a plant DNA sample shows an ratio well below (e.g., ), what contamination is most likely present?

Extraction of genomic DNA from plant by using cTAB method Medium
A. Degraded RNA fragments
B. Proteins only
C. Intact genomic DNA only
D. Carbohydrates, phenols, or residual CTAB

41 In the CTAB extraction buffer, CTAB (cetyltrimethylammonium bromide) is a cationic detergent. At the working temperature of , what is its primary mechanism for enabling DNA purification from plant tissue?

Extraction of genomic DNA from plant by using cTAB method Hard
A. It forms insoluble complexes with polysaccharides and proteins while keeping nucleic acids soluble at high salt concentration
B. It precipitates DNA directly by neutralizing the phosphate backbone charges
C. It selectively binds only to RNA, allowing DNA to remain free in solution
D. It denatures nucleases by lowering the pH of the buffer below

42 A student's CTAB buffer contains M NaCl. If the salt concentration accidentally drops to M during extraction, what is the most likely consequence?

Extraction of genomic DNA from plant by using cTAB method Hard
A. DNA yield increases because low salt favors nucleic acid solubility
B. The chloroform extraction step becomes unnecessary
C. CTAB-nucleic acid complexes precipitate along with polysaccharides, reducing DNA yield and purity
D. Proteins remain fully soluble and are cleanly removed

43 -mercaptoethanol (typically ) is added to CTAB buffer. In plant tissues rich in phenolics, what is its specific protective role?

Extraction of genomic DNA from plant by using cTAB method Hard
A. It chelates ions required by DNases
B. It raises the ionic strength to keep CTAB complexes soluble
C. It lyses the nuclear membrane to release genomic DNA
D. It reduces disulfide bonds and prevents oxidation of phenolics into quinones that bind DNA

44 During chloroform:isoamyl alcohol () extraction, what is the specific function of the isoamyl alcohol component?

Extraction of genomic DNA from plant by using cTAB method Hard
A. It reduces foaming and stabilizes the interphase between aqueous and organic layers
B. It precipitates DNA at the interphase
C. It denatures proteins more effectively than chloroform alone
D. It increases the density of the organic phase to trap polysaccharides

45 After isopropanol precipitation, a student observes a ratio of . What does this most likely indicate?

Extraction of genomic DNA from plant by using cTAB method Hard
A. RNA contamination of the sample
B. Pure double-stranded DNA of high quality
C. Protein or phenolic contamination in the DNA sample
D. Excess salt carried over from the buffer

46 A DNA preparation shows of but of . What is the most probable interpretation?

Extraction of genomic DNA from plant by using cTAB method Hard
A. Heavy protein contamination and no polysaccharides
B. Carbohydrate, CTAB, or salt contamination despite low protein contamination
C. Pure DNA with no contaminants of any kind
D. Degraded DNA sheared into small fragments

47 Why is isopropanol often preferred over ethanol for precipitating genomic DNA in the CTAB protocol?

Extraction of genomic DNA from plant by using cTAB method Hard
A. It precipitates RNA more selectively than DNA
B. Less volume is required ( volumes) so tube capacity and co-precipitation of salts are minimized
C. It requires higher temperatures which improves yield
D. It dissolves polysaccharides better, leaving pure DNA

48 The final DNA pellet is washed with ethanol rather than ethanol. What is the rationale?

Extraction of genomic DNA from plant by using cTAB method Hard
A. ethanol would degrade the phosphodiester backbone
B. ethanol denatures any remaining nucleases irreversibly
C. ethanol dissolves DNA to remove impurities then reprecipitates it
D. ethanol removes residual salts while keeping DNA precipitated and insoluble

49 RNase A treatment is included in the CTAB protocol. At which stage is it most effective and why?

Extraction of genomic DNA from plant by using cTAB method Hard
A. After resuspension of the DNA pellet, incubating at so the enzyme degrades co-purified RNA
B. Before adding CTAB buffer, to protect DNA from RNA interference
C. During chloroform extraction, to partition RNA into the organic phase
D. Immediately after -mercaptoethanol addition to prevent oxidation

50 A researcher grinds fresh leaf tissue in liquid nitrogen before adding CTAB buffer. What is the primary purpose of this step?

Extraction of genomic DNA from plant by using cTAB method Hard
A. It disrupts cell walls and membranes while keeping tissue frozen to inhibit nuclease and phenolase activity
B. It solubilizes polysaccharides directly into the buffer
C. It precipitates genomic DNA as an insoluble complex early on
D. It denatures CTAB so it acts faster on membranes

51 For plant tissues extremely rich in polysaccharides (e.g., some fruits), increasing NaCl to M in CTAB buffer helps because:

Extraction of genomic DNA from plant by using cTAB method Hard
A. Higher salt increases polysaccharide solubility, keeping them from co-precipitating with DNA
B. Higher salt inactivates CTAB, freeing DNA from complexes
C. Higher salt lowers viscosity by degrading polysaccharides enzymatically
D. Higher salt precipitates DNA faster and reduces polysaccharide binding

52 EDTA is a standard component of CTAB buffer (typically mM). Its essential role in preserving DNA integrity is to:

Extraction of genomic DNA from plant by using cTAB method Hard
A. Act as a reducing agent against phenolic oxidation
B. Chelate divalent cations like that are cofactors for DNase activity
C. Buffer the pH to precisely for optimal CTAB action
D. Provide the high ionic strength needed to solubilize CTAB complexes

53 Tris-HCl at pH is used in CTAB buffer. Why is a pH of approximately specifically chosen over acidic pH?

Extraction of genomic DNA from plant by using cTAB method Hard
A. Slightly alkaline pH keeps DNA stable and minimizes depurination and DNase activity
B. pH 8.0 causes polysaccharides to precipitate immediately
C. pH 8.0 maximizes phenol oxidation, clearing contaminants
D. Acidic pH is needed for CTAB to bind DNA, so alkaline pH prevents this

54 On an agarose gel, a CTAB-extracted genomic DNA sample shows a bright high-molecular-weight band plus a low-molecular-weight smear near the dye front. What is the most likely explanation?

Extraction of genomic DNA from plant by using cTAB method Hard
A. Complete degradation of all genomic DNA into oligonucleotides
B. Intact genomic DNA plus contaminating RNA that was not removed by RNase treatment
C. Pure genomic DNA with no contamination
D. Protein contamination trapped in the well

55 If DNA extracted by CTAB fails to amplify in PCR despite good yield, and the ratio is very low, the most probable PCR inhibitor is:

Extraction of genomic DNA from plant by using cTAB method Hard
A. Excess Taq polymerase in the reaction
B. Too little template DNA in the reaction
C. Residual polysaccharides, CTAB, or polyphenols carried over into the eluate
D. Contaminating RNase A enzyme

56 A protocol recommends warming the CTAB buffer to before adding ground tissue and incubating min. Which combination of purposes does this heating serve?

Extraction of genomic DNA from plant by using cTAB method Hard
A. Cooling-induced polysaccharide crystallization and protein renaturation
B. Precipitating DNA and inactivating RNase simultaneously
C. Enhancing CTAB solubility, promoting membrane lysis, and increasing CTAB-contaminant complex formation
D. Activating DNases to remove damaged DNA and denaturing CTAB

57 During phase separation after chloroform extraction, in which layer is the genomic DNA found and why?

Extraction of genomic DNA from plant by using cTAB method Hard
A. Distributed equally between both layers
B. At the interphase, because DNA precipitates with proteins
C. The lower organic layer, because DNA binds chloroform
D. The upper aqueous layer, because DNA is hydrophilic and partitions into water

58 A student obtains a very low DNA yield and notices excessive foaming and an unclear interphase during chloroform extraction. Which procedural error most likely caused this?

Extraction of genomic DNA from plant by using cTAB method Hard
A. Incubating at for too short a time
B. Adding too much EDTA to the CTAB buffer
C. Using liquid nitrogen for grinding the tissue
D. Vortexing too vigorously and omitting isoamyl alcohol from the chloroform mix

59 PVP (polyvinylpyrrolidone) is sometimes added to CTAB buffer for plants like tea or grape. Its specific function is to:

Extraction of genomic DNA from plant by using cTAB method Hard
A. Bind and remove polyphenolic compounds through hydrogen bonding, preventing them from binding DNA
B. Serve as a reducing agent replacing -mercaptoethanol
C. Increase the salt concentration to solubilize polysaccharides
D. Chelate divalent metal ions to inhibit nucleases

60 Given for a diluted DNA sample (dsDNA factor per unit), what is the DNA concentration of the original undiluted stock?

Extraction of genomic DNA from plant by using cTAB method Hard
A.
B.
C.
D.