Objective: DNA is analyzed by agarose gel electrophoresis after being digested with EcoRI restriction endonucleasse. Procedures: λ DNA and puC18 DNA were put into two tubes respectively. Then‚ EcoRI buffer‚ EcoRI enzyme and deionized water would be put into both tubes. EcoRI enzyme was the restriction enzyme that cut the DNA at the specific sequence. The EcoRI buffer enhanced the stability of many enzymes and binds contaminants that may be present in DNA preparations. DI water was used to
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Restriction Enzymes Restriction enzymes (also known as restriction endonucleases) are a group of bacterial enzymes which cut double-stranded DNA (dsDNA) into smaller fragments at specific points. They are a defence mechanism used by bacteria to cleave the DNA of invading viruses‚ thereby restricting their expression. The exploitation of restriction enzymes ability to cut large pieces of DNA into smaller fragments (called restriction fragments) and the highly specific way in which they do this
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determine the unknown DNA plasmid using restriction enzymes and conducting electrophoresis finally comparing the resulting fragments with the known restriction map. In this lab‚ it succeeds in showing the fragments. In this report we will discuss the‚ results‚ limitations and possible errors. Introduction In biology restriction enzymes are used in several ways to modify and manipulate DNA molecules. One common use is to compare pieces of DNA from one that is unknown‚ with fragments of DNA from another source
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Part I – Restriction Enzyme Digestion To begin this experiment‚ the DNA molecules must be cut into smaller fragments with distinct enzymes called Restriction Enzymes through a process called Restriction Enzyme Digestion. Four microtest tubes were labeled 1 through 4 and added 10 µl of Enzyme Reaction Buffer to each of the four reaction tubes using a micropipette. DNA‚ and Enzyme 1 and 2‚ were then added to the reaction tubes using a new micropipette tip for each transfer of DNA and enzyme (refer to
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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 part of the
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Discovery Restriction enzymes were discovered 40 years ago during investigations into the phenomenon of host-specific restriction and modification of bacterial viruses. Restriction enzymes protect bacteria from infections by viruses‚ and it is generally accepted that this is their role in nature. They function as microbial immune systems. When a strain of E. coli lacking a restriction enzyme is infected with a virus‚ most virus particles can initiate a successful infection. When the same strain
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Kenneth Hampton | | |Restriction Enzymes: | |A study in Reactions and Mapping | | | |November 7‚ 2008 | ABSTRACT This experiment will study the
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mechanical and chemical digestion. (a) Ingestion (b) Digestion (c) Absorption (d) Assimilation (e) Egestion TASK 3: The process of digestion involves mechanical and chemical digestion along with the process of: 1. Ingestion 2. Digestion 3. Absorption 4. Assimilation 5. Egestion Using either a starch molecule or a fat molecule explain the journey from the mouth to the anus and indicate clearly points at which mechanical and chemical digestion are taking place:
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Digestion and Enzymes – APP. Hypothesis: The enzyme‚ Amylase which is used to break down carbohydrates will work the best when heated at 40°C. Also‚ as the temperature increases the reaction rate of amylase increases too. However‚ the reaction rate of amylase will start decreasing when the temperature reaches the enzyme’s optimal temperature. Many enzymes are specific for a certain substrate. For example‚ lipase is a specific enzyme for fat substrates and protease‚ a specific enzyme for
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DNA DIGESTION AND ELECTROPHORESIS In this experiment we will be doing a process called as DNA digestion or also known as restriction digest. A restriction digest is a procedure used in molecular biology to prepare DNA for analysis or other processing. It is sometimes termed DNA fragmentation‚ scientists Hartl and Jones describe it this way: This enzymatic technique can be used for cleaving DNA molecules at specific sites‚ ensuring that all DNA fragments that contain a particular sequence have the
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