Introduction Plasmids are circular‚ double stranded extrachromosomal DNA molecules that are found in bacteria which can self-replicate. They are naturally occurring DNA molecules advantageous to the host bacterium by carrying genes which specify metabolic capacities. (Garrett et al.‚ 2010) Besides‚ plasmids exist in a wide variety of sizes from a few thousands to hundreds of thousands of base pairs. Many plasmids have been engineered to serve as plasmids cloning vectors to carry genes. (Synder et
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Title. Restriction Enzyme Mapping of pBR322 Using Agarose Gel Electrophoresis. II. Authors. Author: Partner: Section: Thursday‚ 1:10 pm Date of Experiment: October 25‚ 2012 III. Introduction. Restriction enzymes (or restriction endonucleases)‚ originally isolated from Haemophilus influenzae in 1970‚ are enzymes within a cell that cleave foreign DNA within a specific and predictable nucleotide sequence (known as a restriction site) regardless of the source of such DNA. Such restriction sites
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BIO 219 Group 1 Section 66 October 19‚ 2012 The extraction of purified DNA from A. fischeri by restriction digestion using Sal I enzyme and pGEM for shotgun cloning Introduction: The ultimate goal of this experiment is to isolate the lux operon‚ a targeted piece of DNA that causes bioluminescence‚ from Aliivibrio fischeri and insert it into the DNA of Escherichia coli in order to make it glow. A. fischeri is a gram-negative bacteria which participates in a symbiotic relationship with
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PLASMID DNA ISOLATION‚ RESTRICTION ENZUME DISGESTION AND AGAROSE GEL ELECTRIPHORESIS Abstract: The gel is covered with an ion- containing buffer‚ such as (TAE)‚ that controls the pH of the system and conducts electricity. overall DNA concentration was lower than expected. Using agarose gel electrophoresis is to separate and visualize the DNA fragment‚ which is produced by restriction enzymes . Introduction: The purpose of this experiment is to measure the size of the fragments of DNA and separate
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course. The main focus of the experiment would be how the Restriction Endonucleases cleave the strands of DNA. For this experiment‚ pBR322 was the specimen to use. Restriction Endonucleases work by cleaving the sugar phosphate backbone of specific DNA sites. Restriction enzymes that have been isolated from bacteria have a defensive role. This idea is illustrated when an attacking foreign cell DNA is trying to alter the bacteria; restriction enzymes cleave the DNA rendering it inert. The second
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Purpose: Restriction enzymes cut DNA at a certain palindromic sequence. Three samples of lamda DNA set up to be cut with restriction enzymes PstI‚ EcoRI‚ or HindDIII. There were also two more samples‚ one of these samples was not mixed with any restriction enzyme and the other was a marker‚ which used an enzyme which creates fragments with a known number of base pairs used to create a standard curve. All five samples were put through agarose gel electrophoresis in order to estimate the amount of
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E. Coli Transformation with Plasmid (pGal)‚ pGal Isolation‚ and Analysis of Plasmid DNA Felicia Osadi Bio 22 April 20‚ 2012 Transformation = group 10 Plasmid = group 7 RFLP = group 1 RESULTS Table I. Plasmid Transformation of E. Coli. Plate # | Agar plate | Type | Result | 1 | X-gal | Control | Extensive lawn growth | 2 | Ampr / X-gal | Control | Clear no bacterial growth | 3 | Ampr / X-gal | Transformation | 1 blue colony | Transformation efficiency = 1 transformants
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Isolation of Caffeine from Tea (12A) Introduction When a person drinks tea or coffee‚ it is not pure caffeine. It also has other natural substances. The purpose of this lab was to isolate caffeine from tea leaves. The methylene chloride removed nearly pure caffeine from the basic tea solution. Caffeine is a white‚ crystalline‚ bitter alkaloid‚ C8H10N4O2 ‚ usually derived from coffee or tea. Caffeine is used in medicine chiefly as a nervous system stimulant. Caffeine is used by humans to ward off
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To My parents‚ Mr. Therdsak and Mrs. Aree Jainonthee who always behind every success of my life ACKNOWLEDGEMENT I would like to express my sincere thanks to my thesis advisor‚ Asst. Prof. Dr. Duangporn Pichpol for her invaluable suggestions and encouragement throughout the period of this research. I am most grateful for her teaching and advice on research methodologies and techniques performed in microbiology testing. Her close attention on my working life and my career path are very much appreciated
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Wintersemester / page 1 Experiment 22 Isolation of plasmid-DNA from bacteria and PCR Advisor Konrad Egli: kegli@botinst.unizh.ch Reading Chapters in BBOM 10th: 10.8 BBOM : Madigan M.T.‚ J.M. Martinko and J. Parker: "Brock - Biology of Microorganisms"‚ 10th Edition (2003)‚ Prentice Hall. Objectives Background • Isolation of plasmid-DNA from different bacteria clones • Handling of bacteria clones • PCR-experiment The typical plasmid is a circular double-standed DNA molecule
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