Question

Lab 1 - PCR Pre-lab discussion. Discuss the following among your table prior to beginning the lab. 1. Describe the overall goal of the polymerase chain reaction. 2. List the components that are required for a PCR reaction. 3. A special DNA polymerase called Taq is used in PCR. What is unique about this enzyme and why is it used in the PCR reaction? 4. We will be using genomic DNA as our template DNA in this PCR reaction. How do we determine which segment of the genome will be amplified? 5. Describe what happens at each of the following temperatures during the PCR cycle: 95C: 95C 55C: 72C

Answer 1

1 Goal of the polymerase chain reaction. 50- PCR means polymerase chain reaction o the main puopose or goal of go the ** PCR is to make multiple or enough copies of the target DNA gegion, and 1 t which are then used for the various processes. - multiple copies of DNA arrem pauda are Tag made by using PCR (polymerase chain reachion. 2. The copies of DNA are made into invitro conditions.. LUDEGi Control Omron D oo 2013 27 There are mainly five components are dequired for the per reaction. t. al DNA template. - It I's an sequence which you want to copy. _b] DNA polymerase enzyme - Jt adds nucleonide 1 pairs to the DNA or synthesizes new Dua Strand. c) poimers . Primers it ao short oligonucleotide - sequence. which requires to omplity - the specific region of DNA. dj Mucleotides - four nucleotides ore are required - adenine, guanine, cytosine & thymin el Buffer solution - Buffer is used to provide a stable pH for the reaction and also for enhancing the activity of enzymes.

3] The unique property of this enzyme is in that it is isolated from the bacterium known as Thermus aquaticus and it is an thermophilic bacteria and hence the Tag polymerase is ten e remains stable at very high temperature while Other encimes get denatused at high temperature. na Taq polymerase is an heat stable enzyme - In PcAo reaction Taq polymerase is used for amplification of the DNA f production of new copies of DNA robo aandum Because PCA is generally operated at high temperature reaction & hence per lo requires an heat stable enzyme, and that Tag pole mesaseis an heat stable enzyme hence it's usedlin PCA 47 we can determine the which segmentor the genomic DNA will be amplified is based on primer. Primer is a short oligonucleotide sequence which are used to provide the stasting point for the synthesis of the DNA. Generally the experimenters determines the 8 region that will be copied & according to that segíon the primeros que made. and which determines the al region of DNA that will amplified

Yra 6] g5c - Athis temperature first denaturation of the DNA strand [double strand) ishto single stranded forno occyasi 95€ - DMA is completely becomes single stranded. 550 - Annealing - At this temperature annealing is occurs. At this temperature An primer is anneal to the DNA strand. that means they binds to the complementare site of the target DMA ols and starts geplication n video- 72'c - Pt thi's temperature exten hon is ctivo. Oduos. Or it is also called ees incubation temperature. At this temperature the Taq polymerase starts to extend they primer to form the nascent DNA strand.