PCR Amplification Create millions of copies of the initial DNA PCR Purification Filter out DNA from primers and enzymes Sequencing Prep Run more PCR to make copies of DNA at various lengths DNA Sequencing Sequence machine runs gel electrophoresis to identify nucleotides and determine sequence. Sequence Analysis Computer determines full DNA sequence which can be run and compared. Explain how fluorescent markers help determine a nucleotide sequence. Each nucleotide is assigned
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process of PCR (See Moodle) In laboratories without a high through point of PCR‚ reactions will usually happen using a gel ’agrose’ (See Moodle) it looks like jelly. You can make slabs of this gel. You boil up the agrose which melts‚ then you cast it in a square cast‚ so you have a slab of jelly. You can then put in wells and holes into the slabs. To put in your samples‚ place gel on piece of equipment‚ and then you can run electrical currents through. When you put electric through you have a complete
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Creation of a Bacterial Cell Controlled by a Chemically Synthesized Genome Daniel G. Gibson et al. Science 329‚ 52 (2010); DOI: 10.1126/science.1190719 This copy is for your personal‚ non-commercial use only. Permission to republish or repurpose articles or portions of articles can be obtained by following the guidelines here. The following resources related to this article are available online at www.sciencemag.org (this information is current as of March 24‚ 2013 ): Updated information and
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2520 2500 2000 *light blue is 1 band Experiment The group first designed the materials to create and hold the .75 agarose gel and experiment. Everything except the comb was made of ¼ inch Acrylic Plexiglass. The glass helped the group see what was going on in the experiment. The comb was 3D printed to be more accurate. One material was a casting tray which held the Gel in future steps. The casting tray had to hold at least 50 mL of liquid. They didn’t want the casting tray to hold much more than
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minutes at 72°C. The Amplified DNAs were analyzed by horizontal gel electrophoresis at 100 V in 1.5% agarose gel (wt/v) in 1X TAE buffer (0.04M Tris-Acetate‚ 0.001M EDTA‚ pH 8.0)‚ stained with 2.5X solution of GelRed™ Nucleic Acid Gel Stain (250μL GelRed™ to 1 liter of distilled water)‚ and gels were visualized under UV light and photographed. Afterwards the PCR amplified products were cut out from the gels and purified using the QIAquick Gel Extraction kit (Qiagen). Sequencing reactions were performed
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human insulin? 3. Some people developed allergies to the animal insulin because their immune systems recognized the proteins as foreign. Explain why the immune system would be able to distinguish animal insulin from human insulin. 4. A SDS-PAGE gel is run of proinsulin
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Exercise 13 MITOSIS: REPLICATION OF EUKARYOTIC CELLS ANSWERS TO QUESTIONS 1. a. Mitosis and cytokinesis are often referred to collectively as "cellular division." Why are they more accurately called cellular replication? The result of mitosis is production of two cells (replicates) identical to the parent cell. The genetic material is replicated rather than divided. b. Does the cell cycle have a beginning and an end? The organization of our study of cellular events indicates a beginning and
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“EXPRESSION‚ PURIFICATION AND QUANTIFICATION OF NEURAMINIDASE GENE OF INFLUENZA” A dissertation submitted to the VIT University in partial fulfillment of the requirement for the award of the degree of MASTER OF SCIENCE In BIOTECHNOLOGY SUBMITTED BY A. SRIKANTH Register No: 08MSBOO1 UNIVERSITY (Estd. u/s 3 of UGC Act 1956) VIT SCHOOL OF BIO SCIENCES AND TECHNOLOGY VIT UNIVERSITY VELLORE-632014 MAY 2010 DECLARATION I here by declare that the project work entitled “EXPRESSION‚PURIFICATION
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Expression of a functional recombinant fusion protein via the directional sub-cloning of an E.coli derived tyrosine phosphatase gene (wzb) into a pT5(6H)CFP mutant expression vector. Abstract: Application of fluorescent fusion proteins to the field of expression and interaction proteomics as a means of dynamic imaging proteins in vivo has allowed for rapid advancements in biotechnology research. Production of such proteins first involves the insertion of a given protein-coding gene transcriptionally
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Expressing and Purifying the Recombinant form of Green Fluorescent Protein (rGFP) from the E.coli strain using Ni2+ agarose affinity chromatography technology Abstract The purpose of this experiment was to express and purify the his6-tagged recombinant form of GFP (rGFP) from the organism E.coli using Ni2+ agarose affinity chromatography. The expression of rGFP was confirmed qualitatively using the UV light and was expressed in the E.coli strain BL21 (DE3) (-- removed HTML --) (-- removed HTML
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