BIO203 Midterm Solved Past Papers

Bio203

timing: 10:30 My paper

1. multiplex PCR enables the simultaneous amplification in many targets on interest in ………

One reaction

2. Pcr amplification specificity is ensured by --------(polymerase enzyme)
3. to clone large genes not possible with conventional PCR.( Long pcr)
4. Allele-specific PCR used for identifying of SNPs.
5. In Situ PCR (ISH ) is a polymerase chain reaction that actually takes place inside the cell on a slide.
6. Assembly PCR is the synthesis of long DNA structures by performing PCR on a pool of long

oligonucleotides .

7. In sucide is typically used in polegenetics.
8. PCR used to produce a unique fingerprints of amplified product lengths is called (INTER

SEQUENCE SPECIFIC PCR )

9. RT pcr is widely used in ----( expression profiling).
10. in nested pcr -----used to amplify a fragment.( .two pairs of primer).
11. in molecular technique the RAPD stand for( .Random Amplified Polymeric DNa).
12. Denaturation temperature:Reduces double stranded molecules to single stranded, 90–96oC,

20-45 seconds.

13. primer may occur due to the Hybridization of primer each other.
14. Taq polymerase most commonly used polymerase and code for (Thermus aqauticus)..
15. WGA is teachnique in which amplifications proceed ___in genome.( Directly at any

location)

16. In annealing temp is lowered to ___ for 20 - 40 sec.( 40 - 65)
17. PRIMERS: Primers are single-stranded 18–30 bp long.
18. ...... Controls for contamination contains all reagents except DNA template.(Blank

reaction).

19. Source of Taq polerases the Thermus aquaticus
20. Set of each primers work on the A. high temp

21) What is WGA technique

Answer: Whole Genome Amplification (WGA) is a method for robust amplification of an entire genome, starting with nanogram quantities of DNA and resulting in microgram quantities of amplified products. WGA has become an invaluable method for preserving limited samples of precious stock material, particularly when using WGA methods that have been developed to amplify material from a single cell

Q22. RAP stands for?

23) use of methylation in pcr

Answer: Methylation-specific PCR is used to identify patterns of DNA methylation at CpG islands in genomic DNA. Target DNA is first treated with sodium bisulfite, which converts unmethylated cytosine bases to uracil, which is complementary to adenosine in PCR primers.

� Two amplifications are then carried out on the bisulfite-treated DNA: One primer set anneals to DNA with cytosines (corresponding to methylated cytosine), and the other set anneals to DNA with uracil (corresponding to unmethylated cytosine). MSP used in quantitative PCR provides information about methylation state of a given CpG island.

24) chemical ragents which is use in conventional PCR and also give reson why thy are use

Answer: DNA template � Primers � Buffer � MgCl2 � Deoxynucleotide triphosphates � DNA polymerase

� Distilled water

25) Write the extention rate and source of Taq. Polymerase Vent and Pfu

Answer: 1) Taq pol extension rate 75 Source T. aquaticus

2. Vent extension rate >80 Source Thermococcus Litoralis
3. Pfu extention rate 60 Source Pyrococcus furiosus

26) Differeniate the methodology and principles of Allele specific and AFLP PCR

Answer: AFLP PCR - METHODOLOGY � Genomic DNA is digested with one or more restriction enzymes. tetracutter (MseI) and a hexacutter (EcoRI).

� Ligation of linkers to all restriction fragments. Pre-selective PCR is performed using primers which match the linkers and restriction site specific sequences.

� Electrophoretic separation and amplicons on a gel matrix, followed by visualization of the band pattern. � AFLP is a highly sensitive PCR-based method for detecting polymorphisms in DNA.

� AFLP can be also used for genotyping individuals for a large number of loci.

ALLELE SPECIFIC PCR

� It requires prior knowledge of a DNA sequence, including differences between alleles.

� Uses primers whose 3' ends encompass the SNP.

� PCR amplification under stringent conditions is much less efficient in the presence of a mismatch between template and primer.

This diagnostic or cloning technique is used to identify or utilize singlenucleotide Polymorphisms (SNPs).

Bio203. 28-6-2018. 10:30am

Qno.1.steps of ligation mediated PCR?

Answer: Uses small DNA oligonucleotide 'linkers' (or adaptors) that are first ligated to fragments of the target DNA.

� PCR primers that anneal to the linker sequences are then used to amplify the target fragments.

� DNA sequencing

� Genome walking

� DNA footprinting

Qno.2.uses of methylation specific PCR?

Answer: Answer above

Qno.3.negative control and blank reaction ?

Answer: NEGATIVE CONTROL � Controls for specificity of the amplification reaction contains all reagents and a DNA template lacking the target sequence.