Question

Percentage of denatureation Eppendorf tube Room temperature (7504 90°C Room temperature 3.96% 12 02 45.4%e 77.1% 22 02 56.2% 76.2% 10.5% 3e 0 41.8% 51.8% 27.1% percentage of denatureation percentage(%) room temperature room temperature 75°C 90°C Explain this result. ^ DI

Answer 1

Denaturation is a very crucial process we can observe for proteins (specially for a wide range of enzymes) and also for nucleic acids. Through this process, protein or nucleic acids lose their own structure (secondary, tertiary or quaternary structure) that is present in native form. As we can see in all the three tubes are showing different denaturation in terms of percentage. Initially at room temperature, no denaturation is evident but when temperature increases then rate of denaturation also increases and a maximum denaturation found 90 ^oC followed by 75 ^oC in all the three samples but showing different sensitivity. This is very important step for unwinding double helix  in PCR reaction. So even when temperature reduces from 90 ^oC to room temperature, a considerable amount of denaturation may be evident. Tube 1 showing good denature efficiency at high temperature followed by tube 2 where as in case of tube 3 high temperature showed good denaturation of the product and it continue to denture at lower temperature that is room temperature (~ 24 ^oC ).