UNIVERSITY OF LAGOS

FACULTY OF SCIENCES

DEPARTMENT OF CELL BIOLOGY AND GENETICS

UNDERGRADUATE [B.SC.] ORAL SEMINAR PRESENTATION [CBG 410]

NAME: NLEMADIM, UCHE TONY

MATRIC NO: 150802006

TOPIC: REAL TIME PCR HIGH RESOLUTION MELTING ASSAY.

SUPERVISOR: DR.O.O.IROANYA

DATE: 8th OF MARCH 2O19.

SUMMARY

The polymerase chain reaction [PCR] is a scientific technique in molecular biology to

amplify a single or a few copies of Dna across several orders of magnitude, generating

thousands to millions of copies of a particular DNA sequence [Joshi et al., 2010].

The adoption of the thermostable Taq polymerase in 1988 greatly simplified the process and

enabled the automation of pcr.[Y.m.Dennis Lo et al.,2006]

The real time polymerase chain reaction [pcr], also known as quantitative real-time

polymerase chain reaction [QRT-PCR] or kinetic polymerase [KPCR], is a technique used to

simultaneously quantify and amplify a dna molecule. It is used to determine whether a

specific DNA sequence is present in the sample and if it is present, the number of copies in

the sample [Hongbao Ma et al 2006].

Uses of real-time PCR includes; gene express analysis, diseases diagnosis and management,

high resolution melting curve [hrm] analysis, food testing, animal and plant breeding[Zahra

soheili et al 2014]

High resolution melting [HRM] analysis is a new,simple,powerful and robust method for

detecting dna sequence variants.HRM analysis can discriminate dna sequence based on their

composition ,length ,or strand complementarity[Jesse L Montgomery et al.,2010]

High-resolution melting of dna is a simple solution for genotyping, mutation scanning and

sequence matching [Gudrum H Reed et al., june 2012].

Hrm analysis detects single nucleotide polymorphisms [SNPs] and small insertions or

deletions in a fragment of amplified dna by comparing the fluorescence as a function of the

temperature.[Meistertzheim et al .,2012].

Any double-stranded Dna present will fluoresce strongly at low temperatures, as the

temperature is increased,the fluorescence will decrease at first slowly,and then at a

characteristic temperature the fluorescence rapidly drops,reflecting the melting of dna into

single strand[Gudrum H Reed et al.,june 2012]

Gene or variant scanning is the best- known and most common application of high-resolution

melting[jesse l montgomery et al.,2010]

high-resolution melting analysis is simple,rapid and inexpensive but depends strongly on the

quality of the pcr,instruments and dyes.[ Meistertzheim et al 2012].

In conclusion Real time pcr amplifies the region of dna,for the detection of dna sequence

variance or mutant .

REFERENCES

Dr.Mohini, J. and DR.deshpande ,J.D. 2010.Polymerase chain reaction:methods,principles and

application. international journal of biomedical research review.1[5]:81-97

Dennis Lo,Y.M. Rossa, W.K.C. and Allen chan,K. C.2006. Clinical application of pcr.2nd ed, Hong Kong.
195pp.

Hongbao,M.S. Geroge,C. X.Tracy, Q. and mei-ying, C.2006.Springer,Application of real-time

polymerase chain reaction . The journal of American science review.2 [3]:14-15

Zahra, S. and Shahram,S.2014. National research institute for genetic and biotechnology

Montgomery,J.Sanford, I. and Wittwer, C. 2010. high resolution dna melting analysis in

clinical research and diagnostics.Expert Review of Molecular Diagnostics.10[2].23pp

Gudrum, H.R. Jana ,O.K and Carl, T.W. 2007. high-resolution dna melting analysis for
simple and efficient molecular diagnostics.University of Utah Medical Centre Review.8[6]:12

Meistertzheim, A. Carolyn, F. Sebastien, A. and Christine,P. 2012 .high resolution melting

analysis for last and cheap polymorphism screening of marine population.Research
Gate:31pp