Experiment 2 Titles Extraction of Bacterial Plasmid DNA and Analysis of Extracted DNA Samples Objectives * To learn the procedures needed in extracting the bacterial plasmid DNA * To determine the concentration of original DNA sample and purity of prepared DNA sample by using spectrophotometer * To analyze the extracted DNA sample by gel electrophoresis Materials and methods (Refer to UDBB2144 Laboratory 2A Manual Principles of biotechnology page 6-10) Results
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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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Edwin Southern invented the molecular technique of Southern Blotting in 1975. This technique is very useful because of its ability to detect a specific DNA sequence from a large amount of DNA ⎯ even from the whole genome. This specific sequence can be found through a combination of two different techniques: Agarose Gel Electrophoresis and Hybridization‚ which is known as Southern Blotting. This technique if performed in three phases: (I) prepare the gel and (II) make the blot‚ (III) hybridize and
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AGAROSE GEL ELECTROPHORESIS Gel electrophoresis is a widely used technique for the analysis of nucleic acids and proteins. Most every molecular biology research laboratory routinely uses agarose gel electrophoresis for the preparation and analysis of DNA. We will be using agarose gel electrophoresis to determine the presence and size of PCR products. Background: Electrophoresis is a method of separating substances based on the rate of movement while under the influence of an electric field
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DNA Extraction from Wheat Germ and Making an Agarose Gel AIM: To be able to make and agarose gel and perform gel electrophoresis in six different dyes. Also‚ to extract DNA from wheat germ. INTRODUCTION: Agarose gel is a substance that is used in science for gel electrophoresis and size exclusion chromatography. These processes use agarose gel to separate and analyze proteins and DNA. The medium is composed of a purified agarose powder that has been boiled in a buffer solution and then cooled
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Introduction: 1 Vitamin D is critical for embryonic skeletal formation and long bone growth. Without vitamin D‚ the hypertrophic cell zone will expand because the cartilage fails to calcify leading to rickets. Protein disulfide isomerase A3 (PDIA3)‚ also known as ERp60‚ ERp57‚ Grp58‚ and 1‚25-MARRS‚ is a multifunctional protein that has been associated with rapid membrane-initiated signaling by 1α‚25-dihydroxyvitamin D3 (1α‚25(OH)2D3) in several cell types. Knockdown of PDIA3 in osteoblast-like
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Purification of Infliximab[3] begins at Stage 1 with the filtration of the harvest material using a spin filter. The harvest is then clarified and undergoes 3 main chromatographic steps. Firstly‚ at Stage 2‚ the direct protein capture by Protein A affinity chromatography helps to remove a large majority of the impurities whicn include viruses and media components. At Stage 3‚ prior to cation exchange chromatography‚ the purified material in the eluted stream is then subjected to freezing‚ thawing
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Cell Lab Report Experiments : 1. PCR . 2. Protein extraction and purification . 3. Protein concentration determination . 4. SDS-PAGE . 1. The aim of experiments : 2.1 The aim of PCR experiment is to replicate some DNA dimmers by using specific enzymes used for replication in vitro which is done in lab not by living organisms. 2.2 The aim of protein extraction and purification experiment is to extract some proteins and purify them by specific methods.
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Procedure 2: DNA Extraction from Cheek Cells Materials: Water‚ Clear Dish Soap‚ Table Salt‚ Isopropyl Alcohol (70%) or Ethanol‚ Food Coloring 1. To 200 Ml drinking water add two teaspoons of salt 2. Gargle the salt water for 1 minute. 3. Spit the gargled water into a beaker (or new cup). Now your cheek cells are suspended in the salt water. 4. Gently stir the salt water with one drop of soap (try to avoid air bubbles) 5. In a separate beaker (or cup)‚ mix 20 ml isopropyl alcohol and 1-3 drops
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In Lab 8‚ we had analyzed remains found at a wooded area near Jonesburg and tried to determine if the bones belonged to a 28-year-old woman who had been reported missing from a city within the vicinity. Upon analysis‚ it was determined that they did belong to a female. However‚ it was not possible to determine if the bones did belong to the missing women. Lab 12 presented the opportunity to genetically analyze the remains found. DNA profiling‚ also referred to as typing and fingerprinting‚ uses genetic
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