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The purpose of the PCR (Polymerase Chain Reaction) is to replicate a small strand of DNA sample into a batch large enough to measure through technological means. The DNA sample taken from a patient acts as the template for the replication to base from; meaning after the process of the chain reaction, multiple amounts of the patient's DNA is measurable. While primers in the chemical solution acts as the beginning placeholders for Taq polymerase to add complementary nucleic acids (such as Adenine, Thymine, Cytosine, Guanine) to the DNA template. The deoxyribonucleotides (dNTP's) acts as the building materials in this solution, for the polymerase to attach onto the separated DNA strand. These are the main components for PCR to perform effectively and efficiently. | The purpose of the PCR (Polymerase Chain Reaction) is to replicate a small strand of DNA sample into a batch large enough to measure through technological means. The DNA sample taken from a patient acts as the template for the replication to base from; meaning after the process of the chain reaction, multiple amounts of the patient's DNA is measurable. While primers in the chemical solution acts as the beginning placeholders for Taq polymerase to add complementary nucleic acids (such as Adenine, Thymine, Cytosine, Guanine; binding to Thymine, Adenine, Guanine, Cytosine respectively) to the DNA template. The deoxyribonucleotides (dNTP's) acts as the building materials in this solution, for the polymerase to attach onto the separated DNA strand. These are the main components for PCR to perform effectively and efficiently. | ||
During the PCR process, a cultivating machine heats up the DNA smaple mixture; essentially accelerating the process, which normally proceed slowly without any energy-incentive. The ''initialization step'' of the procedure | During the PCR process, a cultivating machine heats up the DNA smaple mixture; essentially accelerating the process, which normally proceed slowly without any energy-incentive. The ''initialization step'' of the process begins this procedure (368K for 3 min). Next, the ''denaturation step'' separates the DNA template within the mixture into single strands by disrupting the hydrogen bonds between each paired bases (368K for 30 sec). Afterwards, the temperature lowers to 330K (30 sec) to allow primers to ''anneal'' to each patient's individual strands of DNA. This primes the replication process of the PCR procedure. It is by the end of the annealing step that Taq polymerase begins to extend, or ''elongate'', upon each individual DNA strand--pairing free-roaming dNTPs to complementary nucleic bases and synthesizing new pair of the patient's DNA (345K for 30 sec). It is crucial around this time for the DNA strands to form; if the temperature is too high both the primer and the bases cannot bond to their respective region, and if the temperature is too low either may not properly bond. | ||
As soon as the elongation are complete, the PCR cycles multiply times (from denaturating to elongation) until a desired amount of the sample is achieved. From here, the machine proceed with the final elongation (345K for 3 min)--to ensure any remaining individuals to be synthesize into full DNA strands--then cools to 277K for the final product to be used or stored. | |||
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Revision as of 21:42, 29 March 2015
BME 100 Spring 2015 | Home People Lab Write-Up 1 | Lab Write-Up 2 | Lab Write-Up 3 Lab Write-Up 4 | Lab Write-Up 5 | Lab Write-Up 6 Course Logistics For Instructors Photos Wiki Editing Help | |||||||||||||||||||||||||||||||
OUR TEAMLAB 4 WRITE-UPProtocolMaterials
The thermal cycling program is a program that is used to control the OpenPCR machine. Using a varied set of input data, the program can tell the machine to heat up to a specific temperature, cool down to a specific temperature, and specify the number of times to repeat the cycle.
Research and DevelopmentPCR - The Underlying Technology
During the PCR process, a cultivating machine heats up the DNA smaple mixture; essentially accelerating the process, which normally proceed slowly without any energy-incentive. The initialization step of the process begins this procedure (368K for 3 min). Next, the denaturation step separates the DNA template within the mixture into single strands by disrupting the hydrogen bonds between each paired bases (368K for 30 sec). Afterwards, the temperature lowers to 330K (30 sec) to allow primers to anneal to each patient's individual strands of DNA. This primes the replication process of the PCR procedure. It is by the end of the annealing step that Taq polymerase begins to extend, or elongate, upon each individual DNA strand--pairing free-roaming dNTPs to complementary nucleic bases and synthesizing new pair of the patient's DNA (345K for 30 sec). It is crucial around this time for the DNA strands to form; if the temperature is too high both the primer and the bases cannot bond to their respective region, and if the temperature is too low either may not properly bond. As soon as the elongation are complete, the PCR cycles multiply times (from denaturating to elongation) until a desired amount of the sample is achieved. From here, the machine proceed with the final elongation (345K for 3 min)--to ensure any remaining individuals to be synthesize into full DNA strands--then cools to 277K for the final product to be used or stored.
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