HomeCoursesMEDSCI 203: Mechanisms of DiseaseLecture 3 Lecture 3 1. How is DNA selectively amplified? (2) "PCR, cloning"How is DNA selectively amplified? (2)== "PCR, cloning" 2. What is the rate of amplification using PCR? Millions of copies in 2 hoursWhat is the rate of amplification using PCR?== Millions of copies in 2 hours 3. What are the steps in the PCR process? (4)== 1. Template DNA. 2. Denaturation. 3. Annealing. 4. Extension 4. At what temperature is DNA denatured? 95 degrees celciusAt what temperature is DNA denatured? ==95 degrees celcius 5. What happens during annealing? (2) "Temp dropped to 60 degrees, primer added to singular DNA strands"What happens during annealing? (2)== "Temp dropped to 60 degrees, primer added to singular DNA strands" 6. What temperature change allows extension to take place? 70 degrees celsiusWhat temperature change allows extension to take place?== 70 degrees celsius 7. What is extension? Base pairs attached to strandsWhat is extension?== Base pairs attached to strands 8. How many cycles of PCR is the usual standard?= =~30 How many cycles of PCR is the usual standard?= =~30 9. What are the advantages of PCR? (3) "Targeted, allows for analysis of highly degraded DNA samples, can detect sequences not normally present (viruses)"What are the advantages of PCR? (3)== "Targeted, allows for analysis of highly degraded DNA samples, can detect sequences not normally present (viruses)" 10. How are genes separated? Gel electrophoresisHow are genes separated?== Gel electrophoresis 11. What are the different types of gels used for electrophoresis? (3) "Polyacrylamide gels, agarose gels, pulsed field gel"What are the different types of gels used for electrophoresis? (3)== "Polyacrylamide gels, agarose gels, pulsed field gel" 12. How is DNA separated by agarose gel? Agarose gel contains small ‘pores’ which allows smaller DNA molecules to move through it faster than larger onesHow is DNA separated by agarose gel?== Agarose gel contains small ‘pores’ which allows smaller DNA molecules to move through it faster than larger ones 13. What are the advanced methods of sequencing? (2) "Capillary sequencing, Massively parallel sequencing"What are the advanced methods of sequencing? (2)== "Capillary sequencing, Massively parallel sequencing" 14. Which sequencing technique analyses RNA? RT-PCRWhich sequencing technique analyses RNA?== RT-PCR 15. What are the components of a ‘taqman’ probe? (2) "Flourophore, quencher"What are the components of a ‘taqman’ probe? (2)== "Flourophore, quencher" 16. What is hybridisation? Formation of double strand DNA from two single strands with complementary sequences that have be obtained from two sources and denaturedWhat is hybridisation?== Formation of double strand DNA from two single strands with complementary sequences that have be obtained from two sources and denatured 17. What methods are used to denature with hybridisation? "Heat, alkali"What methods are used to denature with hybridisation? =="Heat, alkali" 18. What are some hybridisation methods? (3) "Single nucleotide polymorphism arrays (SNP), microarray, array comparative genomic hybridisation (aCGH)"What are some hybridisation methods? (3)== "Single nucleotide polymorphism arrays (SNP), microarray, array comparative genomic hybridisation (aCGH)" 19. What is RT-PCR used for? Analysing RNAWhat is RT-PCR used for?== Analysing RNA 20. What is aCGH used for?== Detecting gene expression in RNA 21. What are the methods for analysing proteins? (3) "Immunohistochemistry (IHC), western blot, immunoassay"What are the methods for analysing proteins? (3)== "Immunohistochemistry (IHC), western blot, immunoassay" 22. What does an immunoassay tell us about a protein? == Protein amount 23. What does IHC tell us about a protein? (2)== "Amount, location" 24. What does western blotting tell us about a protein? (2) "Amount, size"What does western blotting tell us about a protein? (2)== "Amount, size" Loading...
