ENVB 437 Lecture Notes - Lecture 18: Breastfeeding, Western Blot, Ribosomal Rna

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greywildebeest28

4 Jun 2018

School

McGill University

Department

Environmental Biology

Course

ENVB 437

Professor

Gordon Hickey

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Eukaryotic cells and viruses – lecture 18 (March 21st, 2018)

Slide 4

- Microarrays can be put on chips and use them to screen for diseases and conditions

Slide 5

- Specifically spot a gene sequence that we get with all of our genome sequences

o And we can know the location of all those genes by printing then in a specific location

- By measuring the ratio of the intensities, you can get a measurement of the transcription of this in the entire

cell

→ VIDEO

Slide 6

- Extremely reproducible so if two different labs use these, results should be the same

Slide 7

- After amplification and purification can be printed on a slide with a printer

- You can modify the printer to print on a slide

o Using this technology, you can print easily 10k different probes on a slide

- Each base that is added, is added to a photoprotection group as well

o You can then de-protect different areas

- You apply a new mask and repeat the process over and over again and repeat the process for all the genes

Slide 10

- The major advantages:

o Reproducible – can share protocols

▪ Have confidence in the data that you get

o Since they are so widely distributed and used

- Disadvantages:

o Quality control (QC) is paramount to making good results

▪ They spot mismatch oligos, and if that lights up that means that QC is not very good

▪ The stringency of the washing solution is going to determine which of these sequences remain

present

Slide 11

- You get an idea of the quantity of the different transcripts that are being hybridized

Slide 13

- You have a scale to quantify

Slide 14

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Document Summary

Eukaryotic cells and viruses lecture 18 (march 21st, 2018) Microarrays can be put on chips and use them to screen for diseases and conditions. Specifically spot a gene sequence that we get with all of our genome sequences: and we can know the location of all those genes by printing then in a specific location. By measuring the ratio of the intensities, you can get a measurement of the transcription of this in the entire cell. Extremely reproducible so if two different labs use these, results should be the same. After amplification and purification can be printed on a slide with a printer. You can modify the printer to print on a slide: using this technology, you can print easily 10k different probes on a slide. Each base that is added, is added to a photoprotection group as well: you can then de-protect different areas.

Related Questions

Human body can be described by many means â one of them â it is relatively big vessel with complex body fluid. For diagnostic purpose it is very complicated to detect insignificant number of biomarkers on very early stages of pathological process. This article, present pioneering method to initiate and conduct diagnostic by researching body fluids.

**With additional research provide additional examples of scientific way of dealing with solutions to improve functionality of human life. (QUESTION - This is exactly what is witten on my assignment)**

Analyzing human blood for a very low virus concentration or a sample of water for a bioterrorism agent has always been a time-consuming and difficult process. Researchers at the Georgia Institute of Technology and Emory University have developed an easier and faster method to detect these types of target molecules in liquid samples using highly porous, micron-sized, silica beads. The researchers developed a technique to simultaneously or sequentially add optical and magnetic nanoparticles into the beads. Adding magnetic nanoparticles allows the use of a magnetic field to attract and easily remove the beads from a liquid sample. âThese nanoparticles enter the pores of the microbeads so quickly and so completely -- essentially more than 99 percent of the nanoparticles go into the pores of the beads,â explained Shuming Nie, the head researcher on the project and the Wallace H. Coulter Distinguished Chair in Biomedical Engineering and director of Emory-Georgia Tech Nanotechnology Center. The beads are mixed in a liquid such as urine. Viruses, proteins or other biomarkers are captured on the bead surface. After the beads are removed from the liquid, optical imaging is used to determine the concentration of a specific protein or virus in the liquid sample based on the number of proteins or viruses attached to the surface of the beads. Tushar Sathe, a graduate student in the Wallace H. Coulter Department of Biomedical Engineering at Georgia Tech and Emory University, described the process of creating these novel beads and their clinical applications on Jan. 20 at SPIE Photonics West in San Jose, California. The work was also published in the Aug. 15 issue of Analytical Chemistry. The technology involves embedding fluorescent quantum dots and magnetic iron oxide nanoparticles inside the beads to create dual-modality magneto-optical beads. Nie and Sathe synthesize the quantum dots in different colors by varying their size, giving the beads a unique optical signature. Having different color beads allows the researchers to detect several target molecules at the same time in the same liquid sample. âWe use the quantum dots to create a set of beads that are unique and can be distinguished from each other. Itâs similar to bar-coding -- once you barcode the beads and put them in the urine or blood sample, you can remove them and decode what proteins or viruses have attached to individual beads based on their spectral signature,â explained Sathe. The process of creating these beads is quite simple, according to Sathe. The surface of the beads contains a long-chain carbon molecule that makes the beads hydrophobic, meaning they repel water. The beads are dissolved in butanol and washed several times. Then the beads are counted and optical and magnetic nanocrystals are added to the suspension either simultaneously or sequentially. After 15-20 minutes, the butanol is removed to get rid of any remaining nanoparticles that didnât get incorporated into the beads and the beads are washed with ethanol. Then the beads are coated with a polymer that creates a hydrophilic surface on the beads. This allows the beads to be functionalized by adding antibodies or DNA molecules to the surface that will capture the target molecules. These beads are dual-function -- both optical and magnetic -- but according to Sathe, more functions can be added to the beads. âAdding them is as easy as adding the nanoparticles into the solution. You just have to make sure the nanoparticle surface is hydrophobic so that it interacts with the beads,â said Sathe. The primary biomedical applications for this new technology will be to detect cancer and neurological diseases by identifying certain molecules present in human blood or urine that indicate specific diseases, according to Nie, who is also professor of biomedical engineering, chemistry, materials science & engineering, and hematology and oncology at Emory University and the Georgia Institute of Technology. âSome of the biomarkers for Alzheimerâs disease have very low concentrations in the blood so you need highly sensitive techniques that can find a specific molecule to diagnose this disease,â explained Nie. âOur technique could also be used to monitor therapeutic response. For example, if the viral level decreases in samples taken at later dates, then we know the drug is probably working.â This new technology allows the researchers to analyze very low concentrations of target molecules. âInstead of analyzing a liter of sample where the concentration could be very dilute and you might not see the target molecule youâre looking for, you can let the beads capture the molecules on their surface, remove them from the liquid, and then just measure the number of molecules attached to the beads,â said Nie. This ongoing research is funded by the National Cancer Institute, the Department of Energyâs Genomes to Life (GTL) Program, the Department of Defense and the Georgia Cancer Coalition, a public-private partnership established by the Georgia General Assembly in 2001.

ENVB 437 Lecture Notes - Lecture 18: Breastfeeding, Western Blot, Ribosomal Rna

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