Difference between revisions of "Team:TecCEM/Results"
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| − | <h1 class = "subTitleUbuntu paddingTop">Real Time | + | <h1 class = "subTitleUbuntu paddingTop">Real Time PCR</h1> |
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| − | <p> | + | <p>Since the focus of this project is to verify the effective gene silencing with the four different siRNA we designed, the direct measurement of the amount of mRNA in the cells was required. The first step was to perform a Two-Step Reverse Transcriptase PCR to find out where do the siRNA molecules hybridize with the mRNA. The results are shown below. .</br></br></p> |
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<h1 class = "subTitleUbuntu paddingTop">Diaphorina citri primary cell culture </h1> | <h1 class = "subTitleUbuntu paddingTop">Diaphorina citri primary cell culture </h1> | ||
| − | <p> | + | <p>The main objetive was to develop a primary culture of Diaphorina citri cells, to be transfected with our siRNA and analyzed through flow cytometry. The transfected siRNA sequences were tagged with Alexafluor so they would be visible. This protocol was repeated several times under different conditions. The composition of different culture mediums used is listed below. For the first 12 hours, the cells were able to survive. However, they couldn't replicate and died. Additionally, after 24 hours, a large presence of fungi was observed in the culture. This result was most likely caused by a lack of nutrients in the medium which didn't allow the cells to replicate, as well as a large concentration of serum that allowed for fungi growth. |
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Revision as of 05:22, 1 November 2017
Results
siRNA confirmation
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Sequencing
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Blue BSLA colonies
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Lorem ipsum dolor sit amet, consectetur adipiscing elit. Phasellus efficitur dolor erat, vel lobortis augue mattis nec. Ut sit amet placerat massa. Sed dignissim ante eget nibh sollicitudin, at tincidunt mi fermentum. Curabitur tempus nibh in velit maximus egestas.
Real Time PCR
Since the focus of this project is to verify the effective gene silencing with the four different siRNA we designed, the direct measurement of the amount of mRNA in the cells was required. The first step was to perform a Two-Step Reverse Transcriptase PCR to find out where do the siRNA molecules hybridize with the mRNA. The results are shown below. .
Lorem ipsum dolor sit amet, consectetur adipiscing elit. Phasellus efficitur dolor erat, vel lobortis augue mattis nec. Ut sit amet placerat massa. Sed dignissim ante eget nibh sollicitudin, at tincidunt mi fermentum. Curabitur tempus nibh in velit maximus egestas.
Lorem ipsum dolor sit amet, consectetur adipiscing elit. Phasellus efficitur dolor erat, vel lobortis augue mattis nec. Ut sit amet placerat massa. Sed dignissim ante eget nibh sollicitudin, at tincidunt mi fermentum. Curabitur tempus nibh in velit maximus egestas.
Lorem ipsum dolor sit amet, consectetur adipiscing elit. Phasellus efficitur dolor erat, vel lobortis augue mattis nec. Ut sit amet placerat massa. Sed dignissim ante eget nibh sollicitudin, at tincidunt mi fermentum. Curabitur tempus nibh in velit maximus egestas.
Lorem ipsum dolor sit amet, consectetur adipiscing elit. Phasellus efficitur dolor erat, vel lobortis augue mattis nec. Ut sit amet placerat massa. Sed dignissim ante eget nibh sollicitudin, at tincidunt mi fermentum. Curabitur tempus nibh in velit maximus egestas.
Lorem ipsum dolor sit amet, consectetur adipiscing elit. Phasellus efficitur dolor erat, vel lobortis augue mattis nec. Ut sit amet placerat massa. Sed dignissim ante eget nibh sollicitudin, at tincidunt mi fermentum. Curabitur tempus nibh in velit maximus egestas.
Table: Expected products for the amplifications in the Two-step RT-PCR.
| Product | Expected size |
|---|---|
| Tubulin | 195 bp |
| Rac I | 193 bp |
| WNT | 200 bp |
| AWD | 180 bp |
| SOD | 199 bp |
Lorem ipsum dolor sit amet, consectetur adipiscing elit. Phasellus efficitur dolor erat, vel lobortis augue mattis nec. Ut sit amet placerat massa. Sed dignissim ante eget nibh sollicitudin, at tincidunt mi fermentum. Curabitur tempus nibh in velit maximus egestas. Sed massa ipsum, maximus at dictum dapibus, convallis eget augue. Cras purus mauris, mattis quis ornare a, porttitor non quam. Donec sem felis, feugiat vitae porta sit amet, laoreet a leo. Proin in arcu iaculis, facilisis nisi at, rutrum neque. Nullam condimentum, urna quis pharetra lacinia, justo quam fermentum augue, at porta turpis turpis aliquet risus. Aenean lacinia nunc eu porttitor aliquet. Aenean mattis posuere felis, ac finibus est sodales sit amet. Integer lobortis metus vitae ante sollicitudin pharetra. Quisque egestas sem quis ante tristique cursus. Mauris non blandit velit. Ut euismod ut risus rutrum aliquam.
Diaphorina citri primary cell culture
The main objetive was to develop a primary culture of Diaphorina citri cells, to be transfected with our siRNA and analyzed through flow cytometry. The transfected siRNA sequences were tagged with Alexafluor so they would be visible. This protocol was repeated several times under different conditions. The composition of different culture mediums used is listed below. For the first 12 hours, the cells were able to survive. However, they couldn't replicate and died. Additionally, after 24 hours, a large presence of fungi was observed in the culture. This result was most likely caused by a lack of nutrients in the medium which didn't allow the cells to replicate, as well as a large concentration of serum that allowed for fungi growth.
Lorem ipsum dolor sit amet, consectetur adipiscing elit. Phasellus efficitur dolor erat, vel lobortis augue mattis nec. Ut sit amet placerat massa. Sed dignissim ante eget nibh sollicitudin, at tincidunt mi fermentum. Curabitur tempus nibh in velit maximus egestas.
Lorem ipsum dolor sit amet, consectetur adipiscing elit. Phasellus efficitur dolor erat, vel lobortis augue mattis nec. Ut sit amet placerat massa. Sed dignissim ante eget nibh sollicitudin, at tincidunt mi fermentum. Curabitur tempus nibh in velit maximus egestas.
Lorem ipsum dolor sit amet, consectetur adipiscing elit. Phasellus efficitur dolor erat, vel lobortis augue mattis nec. Ut sit amet placerat massa. Sed dignissim ante eget nibh sollicitudin, at tincidunt mi fermentum. Curabitur tempus nibh in velit maximus egestas.
Lorem ipsum dolor sit amet, consectetur adipiscing elit. Phasellus efficitur dolor erat, vel lobortis augue mattis nec. Ut sit amet placerat massa. Sed dignissim ante eget nibh sollicitudin, at tincidunt mi fermentum. Curabitur tempus nibh in velit maximus egestas. Sed massa ipsum, maximus at dictum dapibus, convallis eget augue. Cras purus mauris, mattis quis ornare a, porttitor non quam. Donec sem felis, feugiat vitae porta sit amet, laoreet a leo. Proin in arcu iaculis, facilisis nisi at, rutrum neque. Nullam condimentum, urna quis pharetra lacinia, justo quam fermentum augue, at porta turpis turpis aliquet risus. Aenean lacinia nunc eu porttitor aliquet. Aenean mattis posuere felis, ac finibus est sodales sit amet. Integer lobortis metus vitae ante sollicitudin pharetra. Quisque egestas sem quis ante tristique cursus. Mauris non blandit velit. Ut euismod ut risus rutrum aliquam.
Lorem ipsum dolor sit amet, consectetur adipiscing elit. Phasellus efficitur dolor erat, vel lobortis augue mattis nec. Ut sit amet placerat massa. Sed dignissim ante eget nibh sollicitudin, at tincidunt mi fermentum. Curabitur tempus nibh in velit maximus egestas.
Lorem ipsum dolor sit amet, consectetur adipiscing elit. Phasellus efficitur dolor erat, vel lobortis augue mattis nec. Ut sit amet placerat massa. Sed dignissim ante eget nibh sollicitudin, at tincidunt mi fermentum. Curabitur tempus nibh in velit maximus egestas.
Lorem ipsum dolor sit amet, consectetur adipiscing elit. Phasellus efficitur dolor erat, vel lobortis augue mattis nec. Ut sit amet placerat massa. Sed dignissim ante eget nibh sollicitudin, at tincidunt mi fermentum. Curabitur tempus nibh in velit maximus egestas.
Diaphorina’s development
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Lorem ipsum dolor sit amet, consectetur adipiscing elit. Phasellus efficitur dolor erat, vel lobortis augue mattis nec. Ut sit amet placerat massa. Sed dignissim ante eget nibh sollicitudin, at tincidunt mi fermentum. Curabitur tempus nibh in velit maximus egestas.
Lorem ipsum dolor sit amet, consectetur adipiscing elit. Phasellus efficitur dolor erat, vel lobortis augue mattis nec. Ut sit amet placerat massa. Sed dignissim ante eget nibh sollicitudin, at tincidunt mi fermentum. Curabitur tempus nibh in velit maximus egestas.
Lorem ipsum dolor sit amet, consectetur adipiscing elit. Phasellus efficitur dolor erat, vel lobortis augue mattis nec. Ut sit amet placerat massa. Sed dignissim ante eget nibh sollicitudin, at tincidunt mi fermentum. Curabitur tempus nibh in velit maximus egestas.
Lorem ipsum dolor sit amet, consectetur adipiscing elit. Phasellus efficitur dolor erat, vel lobortis augue mattis nec. Ut sit amet placerat massa. Sed dignissim ante eget nibh sollicitudin, at tincidunt mi fermentum. Curabitur tempus nibh in velit maximus egestas.
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