19EBT234 Genetic Engineering
(Short Answer Questions)
Unit – 1
1. Explain restriction mapping by taking example.
2. What is DNA polymerase-I larger subunit called as after digesting with
trypsin.
3. Explain different chemicals used in disruption of cell membrane
during DNA isolation.
4. Write about thermostable polymerases used in PCR reaction?
5. Explain different methods used in the identification of purity of
isolated DNA?
6. Explain mechanism of DNA ligase?
7. Explain general characters of cloning vectors?
8. What are the general characters of expression vectors?
9. What are the vectors used for transfer genes into yeast?
10. Explain physical and enzymatic method of disruption of bacterial cell
membrane?
Unit – 2
1. What do you mean by electroblotting used in Western blotting?
2. Explain mechanism of construction of recombinant vector
3. What is a TA cloning?
4. What do you mean by a linker and how it is used in cloning of DNA?
5. What is nitrocellulose membrane and its uses in blotting techniques?
6. What are the different natural methods of gene transfer?
7. If a given eukaryotic gene to be expressed in prokaryote. What is the
promoter to be used and justify your answer?
8. What are dot and slot blots?
9. What are the different types of adopters and their applications?
10. Write about sticky and blunt end cloning of DNA.
Unit – 3
1. Explain on genomic DNA and plasmid DNA of a given bacteria.
2. What is cDNA? How it is useful in expression of eukaryotic genes in
prokaryotes?
3. What is a probe in genetic engineering and its uses?
4. Explain on expression screening of DNA library.
5. How size of DNA library can be calculated?
6. Explain the role of reverse transcriptase enzyme in preparation of
cDNA.
7. Explain the end labelling of the probe DNA.
8. What are the advantages of next generation sequencing techniques?
9. What is a dideoxy nucleotide? How is it used in sequencing?
10. What is a cDNA library?
Unit – 4
1. What is nested PCR and its applications?
2. What is a qPCR?
3. Explain RT-PCR technique?
4. What do you mean by paternity testing?
5. What is a Molecular beacon and its uses?
6. What are the applications of hot start PCR?
7. What is cassette mutation?
8. How is PCR used in induction of site directed mutation?
9. If a PCR is conducted for 25 cycles. How many amplified DNA
fragments are produced?
10. Explain composition of each PCR cycle.
Unit – 5
1. What is a Dicer? Describe its function.
2. What is a RISC and its function?
3. What is a siRNA?
4. What is miRNA?
5. What is Flavr Savr tomato?
6. Applications of antisense RNA.
7. What is a Humulin?
8. What is the source for gene in preparation of Bt cotton?
9. What is TPA? Write its applications.
10. List the negative aspects of genetic engineering.
Essay Questions
(8 Marks questions)
Unit – 1
1. What are the various steps involved in isolation of phage DNA?
2. Explain steps involved in isolation and purification of DNA from E.
coli.
3. What are the general characters of cloning vectors? Explain plasmids
as cloning vector with an example.
4. What are the different types of nucleases and polymerases used in
rDNA technology?
5. Write about nomenclature and classification of restriction
endonucleases?
6. Define restrictions site. How do you estimate approximate number of
restrictions sites present on a given DNA? Types of restrictions sites.
7. Draw the restriction map for the given fragments: EcoRI digest: 1kb,
2.5kb and 9.5kb; Bam HI digest 2kb, 5kb, 6kb; EcoRI+BamHI digest:
0.5kb, 2kb,5.5kb, 4kb and 1kb.
8. What are the characters of Ti and Ri plasmids? How do they are useful
in cloning the genes into plants?
9. Explain different types of topoisomerases and their characters. How
do these are useful in rDNA technology?
10. What are BACs and YACs? Compare and contrast these vectors and
functions.
Unit – 2
1. Write a note on prokaryotic gene expression system.
2. How are linkers and adopters are used in cloning of a gene?
3. What are the different natural gene transfer methods used for
bacteria?
4. Explain the artificial gene transfer methods used in plants and
animals.
5. Write about natural and artificial gene transfer methods for animals?
6. What is the principle of Southern blotting? Write a note on Southern
blotting and its applications.
7. Describe Northern blotting techniques and how it differs from
Southern blotting?
8. How Western botting is performed and explain its applications?
9. How do you express a prokaryotic gene in eukaryote? Explain by
taking an example of Bt cotton?
10. Write a note on In situ hybridisation technique and its application.
Unit – 3
1. What is a gene library? Explain construction of genomic DNA library.
2. What is cDNA? Explain synthesis of cDNA. (CO2 and PO2)
3. Why is cDNA library is usually constructed for eukaryotes? Explain
construction of cDNA library add a note on its applications.
4. Explain screening of library using hybridisation technique.
5. Explain screening of DNA library by expression and antibody-based
methods
6. Discuss different methods used in preparation of labelled probes.
7. Discuss in detail the Maxam-Gilbert method of DNA sequencing.
8. Describe automated methods of DNA sequencing.
9. Discuss in detail the Sanger’s dideoxy method of DNA sequencing with
an example.
10. What are the different types of Next generation sequencing
techniques? Explain any one of the methods in detail.
Unit – 4
1. What is a PCR Reaction? Explain in detail the PCR technique.
2. What is Reverse Transcription PCR and Real time PCR, compare both
the techniques.
3. What is RAPD technique? Explain in detail the conduction of RAPD
and its applications.
4. How do you perform AFLP technique? What are its applications?
5. Discuss in detail the DNA finger printing technique and its uses.
6. Explain different methods of site directed mutagenesis and how they
are useful in rDNA technology?
7. Explain RFLP technique and its applications.
8. How DNA fingerprinting is different from Southern blotting? Explain
technical differences and applications.
9. What are the different variants of PCR? Explain any two variants.
10. What do you mean by molecular markers? Explain the markers used
in DNA fingerprinting and RFLP.
Unit – 5
1. What is RNA interference? Explain its applications in rDNA
technology.
2. How do the recombinant human insulin is produced?
3. Discuss the production of genetically modified Bt Cotton.
4. Explain the limitations and negative aspects of genetic engineering?
5. Discuss various applications of rDNA technology in agriculture.
6. Explain the various applications rDNA technology in Medicine.
7. Discuss applications of rDNA technology in environmental
management?
8. Explain design and application of siRNA.
9. Discuss applications of rDNA technology in Industry.)
10. What are the various achievements of genetic engineering?
