TECHNICAL REPORT ON POLLINATION AND DNA EXTRACTION

40 %
60 %
Information about TECHNICAL REPORT ON POLLINATION AND DNA EXTRACTION
Science

Published on September 26, 2014

Author: cosyshare

Source: slideshare.net

Description

It is great privilege doing my Student Industrial Work Experience with HarvetPlus Nigeria C/O International Institute of Tropical Agriculture (IITA), Ibadan. A huge some of practical knowledge and experience was imbibed in me for a period of six months working with HarvestPlus C/O IITA.
With the mentorship and supervision of Dr. Paul Ilona (HarvestPlusCountry Manager), i was thought the relevance of biochemistry in the research institute. Working under HarvestPlus, I was exposed to different laboratories such as the Virology and Molecular Diagnostic Lab, Tissue Culture Lab, Biochemistry Lab and Microbiology lab.
It’s no doubt that biochemistry as a field of study is best practiced and applied in a research organization like HarvestPlus C/O IITA because the institute is sited in good learning environment with lots of sophisticated equipment and machineries. I was given the opportunity to participate in the on-going research program on biofortification of crops with micronutrients leading to the discovery of Vitamin A Cassava and its retention during processing.
Moreover, I also had the opportunity to work with few Scientists and Research Fellowship Students who shared their various knowledge and experiences with me. I also had the opportunity to exchange knowledge with other biochemistry students from different Institutions also doing their Industrial Work Experience.

A Presentation on Students’ Industrial Work Experience Scheme (SIWES) done at Harvestplus c/o International Institute of Tropical Agriculture (IITA), PMB 5320, Oyo Road, Idi-Oshe, Ibadan, Nigeria Presented By ONYIBA, Cosmos Ifeanyi MOUAU/10/15593 Department of Biochemistry College of Natural and Applied Sciences Micheal Okpara University of Agriculture, Umudike March, 2014 International Institute of Tropical Agriculture – Institut international d’agriculture tropicale – www.iita.org Onyiba Cosmos I. cosmos_onyiba@yahoo.com +2348062633518

Trainings Undergone          Tissue culture Cassava seed production via pollination High quality vitamin A cassava processing Cassava pests and disease control Extraction of Total Nucleic acid (DNA and RNA) from cassava Enzyme-linked Immunosorbent Assay (ELISA) Mechanical sap inoculation of plant viruses Polymerase Chain Reaction (PCR) Gel Electrophoresis International Institute of Tropical Agriculture – Institut international d’agriculture tropicale – www.iita.org

Cassava seed production pollination via Steps involved The male and female flowers of interest are first identified. The female flowers are bagged The male flowers are collected in clean sterile bags fully labelled and kept in cold environment The female flowers are unbagged at mid-day. Pollen is dusted into the stigma. The crosses made is labelled in a tag. The pollinated flowers are bagged and unbagged after 72 hours and allowed to grow into fruits. International Institute of Tropical Agriculture – Institut international d’agriculture tropicale – www.iita.org View slide

Pollination continued…  Fruit develop after 45-72 days. mature fruit turn from dark green to pale.  Mature fruits are collected Into bags, sun dried and allowed to shatter  The Dried Fruits Are Shelled To Obtain Seeds.  Seeds obtained are stored in labelled envelopes and kept in cold environment International Institute of Tropical Agriculture – Institut international d’agriculture tropicale – www.iita.org View slide

TOTAL NUCLEIC ACID EXTRACTION AND PURIFICATION CTAB METHOD CTAB buffer compositionKEY COMPONENTS OF EXTRACTION 20mM Ethylenediaminetetra Acetic Acid. 100mM Trizma Base. 1.4M Sodium Chloride. 0.2% 2- Mercaptolethanol. Sampling: Samples mainly collected are cassava leaves. Lysis: The leaves are grinded in CTAB buffer Removal of Protein and contaminants : Phenol- Chloroform and Isoamyl- alcohol are combined to get rid of proteins and other contaminants. The isoamyl- alcohol prevents foaming effect when phenol denatures protein. Precipitation: Isopropanol stored in -200C is used to aid pellet (DNA/RNA) formation and remove salts from pellet while ethanol is used to further purify the pellet. Recovery: The pellet is then resuspended in sterile water and stored in -200 or used for further analysis e.g PCR, Gel Electrophoresis. International Institute of Tropical Agriculture – Institut international d’agriculture tropicale – www.iita.org

TOTAL NUCLEIC ACID EXTRACTION CTAB METHOD Lysis of cell wall and membrane (Aqueous phase) supernatant containing nucleic acid Centrifugation (Organic phase) containing proteins and other contaminants International Institute of Tropical Agriculture – Institut international d’agriculture tropicale – www.iita.org Removalofproteinand contaminants Chloroform,phenoland isoamylalcohol

TOTAL NUCLEIC ACID EXTRACTION AND PURIFICATION CTAB METHOD DNA Precipitation and Recovery Nucleic acid+ Isopropyl Alcohol Centrifugation Washing with ethanol Refrigeration at -800 C for 15 minutes The pellet is suspended in sterile water DNA/RNA pellet International Institute of Tropical Agriculture – Institut international d’agriculture tropicale – www.iita.org

INSTRUMENTATION The VortexThe shakerThe centrifuge The ELISA Reader and Printer The Gel electrophoresis machine International Institute of Tropical Agriculture – Institut international d’agriculture tropicale – www.iita.org

INSTRUMENTATION The Thermocycler The pH meter The Nanodrop SpectrophotometerThe Gel doc (Biorad) International Institute of Tropical Agriculture – Institut international d’agriculture tropicale – www.iita.org

International Institute of Tropical Agriculture – Institut international d’agriculture tropicale – www.iita.org Onyiba Cosmos I. cosmos_onyiba@yahoo.com +2348062633518

Add a comment

Related presentations

How organisms adapt and survive in different environment.

Aplicación de ANOVA de una vía, modelo efectos fijos, en el problema de una empres...

Teori pemetaan

Teori pemetaan

November 10, 2014

learning how to mapping

Libros: Dra. Elisa Bertha Velázquez Rodríguez

Materi pelatihan gis

Materi pelatihan gis

November 10, 2014

learning GIS

In this talk we describe how the Fourth Paradigm for Data-Intensive Research is pr...

Related pages

TECHNICAL REPORT ON POLLINATION AND DNA EXTRACTION - Science

TECHNICAL REPORT ON POLLINATION AND DNA EXTRACTION ... on Jul 16, 2015. Report Category:
Read more

TECHNICAL REPORT ON POLLINATION AND DNA EXTRACTION - Science

×close share technical report on pollination and dna extraction. embed ...
Read more

Laboratory Report- DNA extraction - Documents

Laboratory Report DNA extraction from wheat germ Name: ... TECHNICAL REPORT ON POLLINATION AND DNA EXTRACTION DNA extraction molecular biology
Read more

Pollen - ResearchGate

We really need to do single pollen DNA extraction for PCR. ... Technical Report: Scotese, ... Dear pollination ecologists,
Read more

Restoring wild bees and crop pollination services in ...

RESTORING WILD BEES AND CROP POLLINATION SERVICES IN AGRICULTURAL LANDSCAPES OF ... Non Technical Summary Bee pollination is ... for DNA extraction.
Read more

CRC for Forestry - Publications - Technical reports

Technical Report 200 ... plantations. 18 pp. Download public report. Technical reports from the CRC for ... throughput DNA extraction for forest ...
Read more

DNA Extraction - SERC

Simply put, DNA Extraction is the removal of deoxyribonucleic acid (DNA) from the cells or viruses in which it normally resides. Extraction of DNA is often ...
Read more

Biotechnology Explorer - UF CPET

Biotechnology Explorer ... For technical service, ... Extraction of DNA From Food Samples ...
Read more