Optimization of PCR parameters towards the development of loop-mediated isothermal amplification (Lamp-PCR) in the detection of fumonisin corn (Zea mays L.) kernels

Date

1-2016

Degree

Bachelor of Science in Agricultural Biotechnology

College

College of Agriculture and Food Science (CAFS)

Adviser/Committee Chair

Eureka Teresa M. Ocampo

Restrictions

Restricted: Not available to the general public. Access is available only after consultation with author/thesis adviser and only to those bound by the confidentiality agreement.

Abstract

To detect and confirm Fusarium and fumonisin gene in Fusarium-infected corn kernels, RNA of grounded and non-grounded corn kernels were isolated using MMY method. These were further subjected to first-strand cDNA synthesis and were run in Polymerase Chain Reaction (PCR) using different primers. Four primers were used to detect fumonisin in general namely Fus 1/2, Fum 5 F/R, Fum 5-5F/6R, PQF19 F/R and Fum 35 F/R while 2 pair of primers which are Vert F-1/2 were for identifying or verifying Fusariumspecies. PCR amplified products from only primer Vert F-1/2 was obtained. Likewise, DNA of the same samples was extracted using CTAB method and was run in PCR using only primer Fus 1/2. All PCR results was run in a 1% gel electrophoresis and then viewed in a Gel Illuminator. The results were sent for sequencing and subjected to BLAST for homology search. BLAST results confirmed and identified that the amplified products from Vert F-1/2 and Fus 1/2 are F. verticillioides intergenic spacer region of the rDNA units and Fusariumfujikuroi IMI 58289 draft genome, chromosome FFUJ_chr04 - a fumonisin gene, respectively. Moreover, Omniamp, a commercially available LAMP-PCR kit was used. Using primer Fus ½ amplification results was obtained. This confirmed that primer Fus 1/2 can detect and amplify fumonisingne in Fusarium-infected corn kernels.

Language

English

Location

UPLB Main Library Special Collections Section (USCS)

Call Number

LG 993.5 2016 A127 /S25

Notes

Major in Crop Biotechnology

Document Type

Thesis

This document is currently not available here.

Share

COinS