Monday, November 9, 2020

RECOMBINANT DNA TECHNOLOGY, CLASS 12, LONG ANSWER QUESTIONS

 3 MARKS QUESTION:

1.     1.  Listed below are the two different ss DNA of their double double stranded form would you expect to be cleaved by a restriction enzyme and why?

(a)    3’ AGCCTTCCGGATGCGC 5’

(b)    5’ GCTTAAGCTTCTGCAAT3’

2.       2. Give three distinguishing features of pBR 322 and pUC 19

3.       3. Write a short note on cosmid vectors and plasmid. Stating the difference between two.

4.       4. What is DNA probe? What is its utility in hybridization technique? Explain it with the help of a hybridization technique.

5.       5. Distinguish between :

(i)                  BAC and YAC

(ii)                M13 and lambda phage

(iii)               Transformation and Transfection

6. Restriction mapping of a linear piece of DNA reveals the following EcoRI restriction sites: 


(a) This piece of DNA is cut by EcoRI , the resulting fragments are separated by gel electrophoresis and the gel is stained . What is the size of the fragments obtained?

(b) If a 1000bp of DNA were inserted between the two restriction sites , what will be the size of fragments when step (a) is repeated.


5 MARKS QUESTION:

 

1.       1. An autoradiogram of a sequencing gel containing 4 lanes of DNA fragments is shown in the

figure:

a.       Describe the method how can this particular sequence be obtained

b.       Describe why the sequence read from the autoradiogram is complementary to the original sequence.


2.    2.   Name the scientist who invented the dideoxynucleotide chain termination method. How would you perform it in single tube? Explain the whole process schematically with diagram for the following strand obtained :

RED -GREEN -BLACK – BLACK -RED- GREEN-BLUE-BLUE-GREEN

Where Red codes for ddA, Black for ddT, Green for ddC, Blue for ddG

3.       3. Describe the principle and use of blue white selection of rDNA technology. Name any two methods of introducing recombinant DNA into host cells.

4.      4.  Explain the function of any three enzymes used in RDT experiments. What is the significance of RM system?

5.       5. Explain with suitable diagram, the steps and principle involved in Sanger’s method of DNA sequencing. 

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