Submitted toSubmitted by- Harshita Jain4th yr B.Tech

Introduction Reaction Components Applications in agricultureConclusionReference

The POLYMERASE CHAIN REACTION (PCR) is a biochemical technology to amplify a single or a few copies of a piece of DNA across several orders of magnitude, generating thousands to millions of copies of a particular DNA Sequence.

It was invented in 1983 by Dr. Kary Mullis, for which he received the Nobel Prize in Chemistry in 1993.

Target DNA contains the sequence to be amplified.Pairs of Primers oligonucleotides that defines the sequence to be amplified.dNTPs deoxynucleotidetriphosphates: DNA building block.Thermostable DNA Polymerase enzyme that catalyzes the reaction. Mg2+ ions cofactor of the enzyme.Buffer Solution maintains pH & ionic strength of the reaction solution suitable for the activity of the enzyme.

Product development Grain processing Identification of Fishery products Cultivar Identification of rice Quantification of Fusarium culmorum in Wheat

and Barley

Major use of PCR technology in product development includes-

Gene discovery and cloning. Vector contruction Transformant identification Screening and characterization Seed quality control.

I. PCR testing for unapproved events

II. PCR testing for gm content

III. PCR testing for non gm labeling

IV. PCR testing for presence of high-value commodity

The importing country often requires that the grain shipment be tested for the presence of specific GM events to ensure that the grain shipment does not contain these unapproved events.

Such testing often relies on qualitative PCR.

Most countries that have adopted mandatory labeling rules for food or feed have set tolerances for the adventitious presence of GM material in grain products.

To meet this need for testing, several laboratories currently are adopting quantitative PCR for percent GM determinations.

In some cases, food manufacturers and retailers wish to use positive labeling for their non-GM products.

The use of positive labeling requires that the grain and grain products originate from a non-GM identity preservation program and test negative or at least below a certain threshold for GM DNA.

Qualitative PCR testing is most often used to certify compliance with a non-GM contract.

In certain cases, it is desirable to show that a commodity is made up of a specific crop commodity

e.g., low phytate maize, soybean with altered oil profile.

PCR could be used for this purpose by testing for the GM trait that conveys the characteristic, although the grain may also be tested by quantifying the improved quality of the commodity.

A method of DNA analysis has been developed to verify authenticity of labelled raw material of canned Fish or in products made from closely related Fish species (tuna, eel, salmon, trout and sturgeon).

Short segments (358 bp) of the mitochondrial cytochrome b gene were amplified by the PCR to get species-specificity patterns.

Cultivars of rice markedly affect eating quality, processing suitability, and price, identification or differentiation.

It became possible to differentiate 60 Japanese dominant rice cultivars from each other using template DNA extracted and purified from rice grains. 

Specific detection of Fusarium culmorum in infected seeds

The specificity of the assay was confirmed by test in seven Fusarium species and 21 non-Fusarium fungal species.

 Eight barley and nine wheat varieties infected by F. culmorum isolate were evaluated in 1 yr (barley samples) and in 4 yrs (wheat samples). 

PCR technology is often used for the detection of products of agricultural biotechnology.

PCR has revolutionised molecular biology and made PCR the most widely used and powerful technique with great spectrum of research applications.

T.A.Brown, DNA Cloning & Gene Analysis.

http://onlinelibrary.wiley.com/doi/10.1002/(SICI)1097-0010(199705)74:1%3C35::AID-JSFA765%3E3.0.CO;2-2/abstract;jsessionid=6B4E317EC6C9F5A29679B9E9E0B86002.f04t04http://pubs.acs.org/doi/abs/10.1021/jf062737z