SYY.BSe. Biotechnology Semester IV (Revised) Examination Biotechnology- Molecular diagnostics Model Answers Do as directed (Any fiieen) 6 Tie Tre Th Southern bloting, the doublestranded DNA Ragments in the gel must ‘be denatured and transferred to a nitrocellulose membrane ‘Restriction endonucleases are also called as restriction enzymes Which recognizes a specific nucleotide-pair sequence in DNA called restriction site and cleaves the DNA (bydrolyzes the phosphodiester backbones) within or near that sequence. Fill inthe blank: Protein ae scparsted using SDS-polaersiamide ae (SDS-PAGE or itoletric focusing gel EF) Fill n the blank: Taq DNA polymerase enzyme fom Thermus aguaticus has facilitated the process of molecular disgnostis, In patemity testing, iT man lacked the allele that must ave come from the baby’s father, then the DNA typing data would have proved that he is ot the father; this is known as Exclusion result Give an example of chelator present in master mix EDTA, The type of PCR coniral which ensures that the enzyme is active, he buffer is optimal, the primers ate priming the write quences, and the thermal cycler is cycling appropriately is called as Positive control —_ used fo increase the effectiveness oF UW Tight to decontaminate and maintain pre-PCR area. Psoralens ‘The Tih polymerase is isolated from Thermus thermophilus Ka ive an example of Monovalent cations present in PCR master mixture | | Define te tem “amplicons” ‘An amplicon is a piece of DNA or RNA that is the source and/or product of amplification. ‘What is the significance of termocyelers? ‘Thermocyclers are designed to rapidly and automatically ramp (change) through the required incubation temperatures, holding at ach one for designated periods. Presence of quasi species of FTV is due to the ror prone nature of he reverse transcriptase enzyme. Stat tre or false True 16 Tes the time period between exposure to HIV and Before seroconversion |isknown as Window periodPace-7is a second generation hybridisation asays used Yor Negonorrhocae. Sate tue oF false False ____ Bene analysis s done to dtect agile X diagnostic. FMRI gene 1. Intrapartum transmission of N gonorrhoeae produses condition in neonates Conjunctivitis oF Ophthalmia neonatorum the process of determining an individual's DNA characteristics, which are as unique as fingerprints. DNA profiling DNA fiagerprintin I QA ‘What is molecular diagnostics? Explain in detail the of mutation detection techniques in molecular diagnostics Definition (IMark) ignificance of PCR (2 Marks) ‘Techniques developed - Enzymatic based methods, Electrophorectic based method, Solid phase based techniques (Marks oF iaborate on FISH asa hybridisation technique FISH: Full form 1 mare ‘Technique and Applications ~ 6 marks ‘OR are DDisauss the organic and inorganic method for olation of DNA Organic DNA isolation method ~ Diagram (1 Mark) Steps and significance of phenol chloroform mixture (3 Marks) Inorganic DNA isolation method — Diagram (I Mark) Salling out method (3Marks) QD Explain westem bloting in detail, The immobilized target for a Wester bot is protein (IMark) Separation using SDS-polyacrylamide gels (SDS-PAGE) or isoelectric focusing gels (TEF) (Mark) Significance of chemicals used and blocking(3Mark) Disadvantage of using denaturing chemicals (Mark) Membrane used (Mark) 7 [Osa Give working principle oF gRT PCR, Principle 2M Steps involved- 3M. ‘Analysis of result- IM Diagram-IM_ O38 Discuss any three components of PCR Principle of PCR-IM Any three component: 2M each ‘© Template = Primer‘+ Deoxyribonucleotde Bases + Thermocycler OR aac ‘What iste finetion of DNA payaeras a PCR What are the ize ‘ypes of DNA polymerases usd in PCR method? unction- 2M Types eM. + Tag polymerase Tin polymerase Stoel agent © ThermoSequenase + T7Sequerase a3D ow io eoairol contannation ia PCRT Significance of contamination-IM ‘Methods | Physical method-2M Chemical method-2M Use of enzymo-2M. o QA ‘Wiite a note on uses of genetic testing Introduction IM Uses: IM each Diagnostic testing Carer testing # Predictive = Suscepi ‘Conclusion - IM lity genetic testing 04D | Write a note on HIV and clinical significance of viral Toad i the serum sample? Introduction- 1M HIV strucure-2M Virology 2M Correlation of viral load with prognosis ofthe disease: 2M. ‘OR Qc How RFLP is used to detec sche call anaemia? Introduction about sickle cell anaemia (Mutation)-2M [RELP principe IM Detection of sickle cell anaemia using RFLP. 3M Diageam-1M. Qa ‘Write a note on clinical significance of N gonorrhoeae? IM foreach point ‘© Common sexually tansmitted microbe ‘Transmission requires direct, usually sexual contact ‘© Iteanbe symptomatic or asymptomatic ‘Intrapartum transmission can also occur, producing disease in eonates 7