In your own words, describe why PCR was important to our lab objective. Material
ID: 174382 • Letter: I
Question
In your own words, describe why PCR was important to our lab objective. Materials and Methods What is the purpose of the Taq DNA polymerase? What is the purpose of the INSTA-GENE? Why are we performing two PCR reactions (PMM and CMM) on each sample (test, non, pos)? Results. Attach a copy of your gel with this worksheet. On the gel: circle any bands which deviate from the expected results. draw in bands that are missing, but should be there if your gel shows ideal results, provide a copy with no marks write a figure legend for your gel in which you: state what your test food was and whether or not is was modified. Make sure to refer to the bands that support your conclusion. If the gel is inconclusive, why is it inconclusive and provide an explanation for what may have gone wrong. make sure to include any additional essential information in your legend. we expect this figure legend to be about a medium-sized paragraph in lengthExplanation / Answer
Q1.WHY PCR WAS IMPORTANT.......??
ANS-Polymerase Chain Reactio(PCR) is one of the most scientfic advances in the field of molecular biology.Through PCR ,it is possible togenerate milions of DNA copies from a single strand of DNA.it has many applications----
MEDICAL--I)Used to anaslyze presence of genetic disease mutations.2)tissu typing prior to organ transplantation
FORENSIC-1)Fingerprinting:used to incriminate or rule out suspects in a crime investigation.2)Parental Testing:used to confirm the biological parents and also to identify the remains of an unidentified body.
MOLECULAR BIOLOGY-1)DNA sequencing,DNA clonning,2)production of hybridization probes for both northern and southern blooting hybridization.
Q2A.WHAT IS THE POURPOSE OF Taq DNA polymerase?
ANS-Taq DNA polymerase is found in bacteria that live in Thermophilic conditions(DERIVED FROM Thermus aquaticus))
It is used to synthesize a new DNA strand from a template inj PCR. During PCR ,the DNA is subjected to not so plesant process like denaturation by heating at high temp. and annealing.Enzymes being protein in nature can be easily denatured which means that they wont be able to perform the required function.
Its speciality is not to get denatured and function effetively even at high temparature.
Q2B:WHAT IS THE PURPOSE OF INSTA GENE?
ANS--INSTA GENE is nothing but the buffered versiopn of the chelating ion erxchange resin chelex-100 and the insta gene matrix solutionn is negatively charged(-ve) microscopic beads which can bind divalent cations.
The insta Gene matrix eliminates the time consuming and labour intensive deproteiniztion,organic extraction,dialysis and alcohol ppt prorocols required in traditional DNA purification process.
Place the cells in microcentrifuge tube,adda the insta Gene matrix ,boil and spin.The matrix adsorbs cell lysis products that interfere with the PCR amplication process.DNA in the supernatent is ready for PCR.
Q3--WHY ARE WE PERFORMING TWO PCR REACTIONS ON EACH SAMPLE?
ANS-It is always goodto run duplicates to make sure the primers are flanking the sequence of interest.
REST QUESTIONS(4-1,2,3,4,5,6) ARE BESED ON EXAMINERS RESULT IN LAB.