thermocycler.pptx

thermocycler

1. THERMOCYCLER Submitted by , Moksha T Jr.M.Sc.(Hort.) Vegetable science COH, Bengaluru UNIVERSITY OF HORTICULTURAL SCIENCES, BAGALKOT COLLEGE OF HORTICULTURE, BENGALURU Submitted to , Dr. Shankarappa K S Assistant Professor Dept. of Plant Pathology. COH, Bengaluru

2. Introduction PCR machine is also known as thermal cyclers or DNA amplifiers A PCR machine is an instrument that amplifies target nucleic acid sequences into millions of copies via polymerase chain reaction Thermal cyclers inherit their name because they regulate temperatures in a cyclical program G- Storm GS4 Thermal cycler system

3. In 1987, the first commercial thermal cycler, was TC1 DNA Thermal Cycler designed by Perkin Elmer Cetus ( American Global Corporation) Principle : “the basic technological principle of heating block is the Peltier effect . By reversing the direction of current between two semiconductors , the heating and cooling phases are alternated . A PCR thermal cycler can thus very quickly reach temperatures between 4 o C and 96 o C required for the PCR”. Principle of Thermocycler TC1 DNA Thermal cycler

4. Parts of a PCR machine

5. How does a thermocycler work? A thermal cycler is a programmable apparatus that works in order to hold materials at different temperatures It consists of a heating block that has depressions or holes so as to hold sample tubes or sample vessels When the apparatus is programmed for PCR, it operates at specific temperatures for specific amount of time The programming can be done by the users with the help of a console on the thermal cycler or a computer Sometimes the instrument comes with a built-in pre-programmed routine

6. Role of the thermocycler in PCR It is the most important technology for scientists who want to make use of polymerase chain reaction (PCR) Thermocycler tightly controls the fluctuation in the temperature needed for annealing, denaturation, and extension It also regulates the number of cycles The thermocyclers which are used nowadays are fully programmable and enable rapid cooling and heating and hence the tighter control of the PCR

7. Types of thermocycler Real time thermocycler Digital thermocycler Fluorescence thermocycler Peltier effect thermocycler Microfluidic thermocycler LAMP thermocycler

8. Applications Carrying out the polymerase chain reaction of DNA for scientific experiments Helping in maintaining the temperature during biochemical reactions such as DNA cloning for sequencing, DNA amplifying, and DNA-based phylogeny Heating and cooling the DNA samples for genotyping Carrying out genetic tests that are helpful in clinical diagnosis of certain diseases Creating thermal gradients in order to find the optimal PCR conditions in experiments like primer annealing

9. Precautions while using PCR Machine Only tubes and plates designed for the PCR machine should be used Before running, double-check tubes and, in particular, plates are tightly sealed Solution spills should be cleaned up, and biohazard containers should be used for disposal Be cautious when using a PCR machine lid . If you drop or bang lids, this could get broken Once, the use of the PCR machine is finished, it should be turned off Prior to beginning a run, make sure the PCR heating block is clean . Before beginning, inspect every tube holder E nsure that the lid sits flat against the top of the tubes for even heating and sealing, distribute the tubers evenly over the block

10. Examples of PCR machines Biometra TAdvanced Thermal Cycler Series (Analytik Jena) MiniAmp™ Plus Thermal Cycler (ThermoFisher Scientific) PCR Thermal Cyclers (Esco) GET-S series thermal cycler (Bio- gener )