Research On The Expressed Sequence Tag Of Olives

835 words - 4 pages

Introduction
Olive, or scientific name Olea europaea L which belongs to Oleacea family is a valuable plant species where it can produce oil and table olives. Gene at certain tissue in species will be expressed where cDNA are sequenced to produce expressed sequence tag (EST) library. EST database is beneficial as it allows new gene discovery, marker discovery, gene mapping, and functional studies to be carried out. This research has contributed EST library of 2304 clone sequences from the young olive leaf and 1536 clone sequences from the immature olive fruit for Turkish olive cultivar Gemlik. Good quality ESTs are used to further analysed by using Phred-Phrap and Contig Assembly Program 3 (CAP3) software. They were then submitted to GenBank (dbEST). BLAST and BLAST2GO are conducted to perform annotation of the sequences.

Methodology
Total RNA was isolated from fresh young leaves and immature olive fruits with RNeasy PlantMiniprep kit (Qiagen). Total RNA was then converted into 1.5 μg and 3μg mRNA respectively by purification, using the Oligotex mRNA Mini Kit (Qiagen). DNA libraries were then constructed with the CloneMiner cDNA Library Construction Kit (Invitrogen). cDNA was cloned into pDONR222 vector and transformed into E.coli strain DH5 (Invitrogen). The cDNA library was plated onto LB-kanamycin agar medium. Colonies which were grown individually were then picked into 384-well plates with SOB medium to be inoculated overnight. They were then added with glycerol. The library was stored at −80˚C. The above steps were done separately for the two cDNA library.
Sixty clones randomly selected. Alkaline lysis method was used to isolate plasmid DNA from the clones selected previously. Bgl1701 was then used to digest the DNA.
Polymerase chain reaction (PCR) was carried out to amplify the clones to create library. 3840 clones were selected randomly to be used as PCR template. M13 universal primer was used as primer in PCR. Automated sequencing was performed using the ABI 3730 capillary sequencer (PE Applied Biosystems) to obtain the sequences of clones.
Low-quality EST sequences were removed by using Phred software. EST sequences with vector sequence were edited using Phrap “cross-match” application. Contig Assembly Program 3 (Cap3) was used to assemble the sequences obtained from sequencing for analysis while Consed/Autofinish software was used to control the sequence assembly. All sequences were assembled separately into contigs. BLAST of sequences was conducted to determine the gene homology in order to connect their functions. Unique sequences were analysed for biological characteristics as well as functional annotation using program BLAST2GO. New genes can then be...

Find Another Essay On Research on the Expressed Sequence Tag of Olives

The Sense of Sadness and Loss Expressed in the Two Poems, On My First Sonne and Refugee Mother and Child

1058 words - 4 pages way Chinua Achebe tries to bring about sadness so does Ben Johnson as they both watch what they have lost or are about to loose. ‘On my first sonne’ and ‘Refugee mother and child’ both have many biblical allusions and the first poem refers to the relationship between Jesus and God because the child was Ben Jonson’s first born just like Jesus and God. In the first stanza Chinua Achebe says “no Madonna and Child could touch that picture of a

Putting a Price Tag on Information: A Study of Electronic Data Interchange

816 words - 3 pages Information technology (IT) can be viewed as a financial drain on a company since it is a huge investment that requires justification. Previous studies have not been able to accurately return information that would lead to a valuable return on investment (ROI) because of the complexity of the technology. The necessary factors had not been taken into account. Factors like the quality of the design of a system and the length of time data had been

The Impact of GPA, Multiple Clinical Rotations, and the Sequence of the Clinical Rotation on Graduates' of College of Pharmacy Performance on the NAPL

1513 words - 6 pages Review Article The aim of this study is to determine the impact of Grade Point Average (GPA), multiple clinical rotations, and the sequence of the clinical rotation on graduates’ of college of pharmacy performance on the North American Pharmacy Licensure Examination (NAPLEX test). Clinical rotations are defined as those clerkship experiences that are not based solely on order entering, or dispensing of medication but requires pharmacist

Analysis of the Opening Sequence of the Simpsons

2107 words - 8 pages Analysis of the Opening Sequence of the Simpsons Matt Groening changed television forever when he brought animation back to primetime with this immortal nuclear family. It was first screened on 14th January 1990 and it has become one of the world’s most famous cartoon as it always captured a regular audience of 24 million each week. It has also established itself as an award-winning international pop culture

A Textual Analysis of the Opening Sequence of Gladiator

1849 words - 7 pages A Textual Analysis of the Opening Sequence of Gladiator In this essay, I will explain the opening sequence of Gladiator in detail. I will describe the effects it has on the audience, and look at the way it makes them feel and the way in which events are portrayed. I will look at in depth: The themes and atmosphere, the camera

A Review of the Opening Sequence of Pretty Woman

2352 words - 9 pages A Review of the Opening Sequence of Pretty Woman Pretty Woman is a romantic comedy, which is based on the stories of Cinderella, the Prince and the Pauper and My fair lady. The film was produced in 1990 and directed by Garry Marshall. It is set in the Hollywood hills and the rough Hollywood Boulevard. Richard Gere and Julia Roberts play the two main characters, Vivien Ward and Edward Lewis. Edward is a wealthy man who

An Analysis Of The Energizer Bunny Commercial Sequence

743 words - 3 pages company was awarded an Obie (the advertising equivalent of the "Oscar") as best commercial of the year. This essay shall attempt to analyze the series of "Energizer bunny" advertisements. There is a current trend in modern television advertising for a series of commercials for the same product. An excellent example is the ad sequence for "Taster's Choice" coffee brand, where a man and a woman share (cups of

How Love is Expressed in the Hunchback of Notre Dame

1151 words - 5 pages Throughout the novel, The Hunchback of Notre Dame, many forms of love appear. There is true love that comes from the heart and is pure and on the other hand there is a lustful love which is not love at. Another love that is shown throughout the book, is love obsession, where one falls in love with someone and puts them before everything. In the Hunchback one can observe these loves and how they compare to one another as well as which one is best

Ethical views expressed in 'Lord of the Flies'

1085 words - 4 pages The Ethical Views Expressed In Lord Of The Flies.In this human world, wherever we turn there will always be conflict. No human isperfect, and all humans err. Therefore we should understand that to err is only human, and toforgive is divine. The ethical views expressed in the renowned bestseller novel, Lord of theFlies, brings into perspective the ever changing external and internal conflict that lies withineach of us.None of us can quite

Christian Culture Expressed in the Music of Hillsong United

1461 words - 6 pages the Christian worldview in Hillsong United’s anthology is the repeated theme of individual redemption. An example of this theme is found in the band’s song “Lead Me to the Cross.” During the anthem, lead vocalist Brooke Fraser declares, “Lead me to the cross, where your love poured out. Bring me to my knees, Lord I lay me down. Rid me of myself, I belong to you” (2007). The contemplative words are a cry of personal reliance on God’s grace and

The Different Types of Love Expressed by Pre-1914 Poets

792 words - 3 pages The Different Types of Love Expressed by Pre-1914 Poets These 3 poems are completely different to each other. Porphyria’s Lover is obsessive and violent love, How Do I Love Thee? Is more of an undying love and the poem Remember is more like truthful love. In this essay I’ll take each poem and in a sentence or two explain the type of love that each poem demonstrates. I’ll also use quotations to show each of my poems shows a different

Similar Essays

Essay On The Idea Of Marginality Expressed By Bell Hooks

929 words - 4 pages live and work outside society's center, and take up space in the margins, much like the margins of a sheet of paper. Author bell hooks, however, believes these margins not only represent sites of oppression, but also serve as places of resistance. To hook's, the margins are something the marginalized need to hold on to, for it is a site of resistance that allows the oppressed to hold on to their individuality and use it as a source of

Analysis Of The Opening Sequence Of Clueless

1237 words - 5 pages Analysis of the Opening Sequence of Clueless The film “Clueless”, written and directed by Amy Heckerling in 1995, is an adaptation of Jane Austen’s early 19th century novel “Emma”. In order to translate the insular world of the provincial English town into a modern scenario, Heckerling hit upon the modern American high school, with its cliques and rituals. Although it appeals to a larger audience, the target audience is

Sequence Of Events: The Cuban Missile Crisis

646 words - 3 pages , 1960, he openly stated that he agreed with the Soviet Union and their policies (Cuban Missile Crisis Timeline). This enraged the United States, as they were already in a conflict of whether capitalism or communism was better with the Soviet Union. On January 3, 1961, the United States terminated all diplomatic relations with Cuba (Cuban Missile Crisis Timeline). Then on April 17, the United States launched the Bay of Pigs Invasion, where a group

The Implications Of Expressed Emotion In Schizophrenia

1936 words - 8 pages , expressed emotion plays a important role in the relapse of Schizophrenics in convalescence (Vaughn and Leff, 1976). Vaughn and Leff also carried out a study which concentrated on the significance of expressed emotion and how this affects the recovery of Schizophrenics. Home environments where there is a low level of Expressed Emotion were shown to have low rates of relapse, compared to high expressed emotion families. When medication was