chromatography * 8.4.3 Purification of a tagged protein * 8.5 HPLC * 9 Concentration of the purified protein * 9.1 Lyophilization * 9.2 Ultrafiltration * 10 Analytical * 10.1 Denaturing-Condition Electrophoresis * 10.2 Non-Denaturing-Condition Electrophoresis * 11 References * 12 External links | Purpose Purification may be preparative or analytical. Preparative purifications aim to produce a relatively large quantity of purified proteins for subsequent use. Examples
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insertion mutations‚ recombination between elements‚ gene conversion‚ and alterations in gene expression. In the lab PCR was used to amplify a short piece of DNA from human genome which allowed us to look for a DNA sequence called an Alu element. Electrophoresis was used to separate DNA fragments of different sizes. The data indicated that the most individuals had the Alu element TPA-25. According to the results x2 was 1.842 because number of homozygous AA individuals without Alu insertion was 5‚ number
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Isolated DNA Products Amplified Via Polymerase Chain Reaction and Cloned Biotechnology: DNA WPUNJ December‚ 2012 Abstract Isolated DNA from mouse‚ plants‚ and plasmid DNA were used for Polymerase Chain Reaction (PCR) for DNA amplification. The purpose of this experiment was to study the success rate or optimization of PCR of DNA‚ using both manual and kit methods. This set of experiments gives an insight to the relative difficulties associated with the optimization of a variety
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This allows an equal concentration of protein from each sample to be load into the wells later on when the gel electrophoresis is run. The Coomassie dye is brownish/red in color but turns blue when mixed with proteins. The blue color can be measured in a spectrophotometer at 595nm. A standard curve was generated using known amounts of bovine serum albumin (BSA). Four
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humans and can be used as a way to distinguish between males and females. Gender determination of a human has important applications in forensic science‚ prenatal diagnosis‚ etc. This can be done by Polymerase Chain Reaction (PCR) followed by gel electrophoresis. The presence of two bands of DNA indicates that the sample is from a male while the presence of one band of DNA indicates that the sample is from a female (sasaki and shimokawa‚ 1995). In this experiment‚ Chelex was used to extract nucleic
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Gel electrophoresis was carried out according to buffer system discussed by [11]. Complisation of electrophoresis gel was processed for activity of proteases detection by gel X-ray film contact print method (GXCP) [12]. After electrophoresis gel was incubated in 0.1M Glycine-NaOH buffer pH 9 for 7 to 8min then was placed on X-ray film [Padul‚ M.V. et al.‚ 2012]. The gel was removed from X-ray film after 45min depending on the extent
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The Expression and Purification of Recombinant Green Fluorescent Protein (rGFP) From E. coli strain‚ BL21(DE3)‚ Using Ni2+-Agarose Affinity Chromatography Abstract: The purpose of these series of experiments was to express and purify recombinant Green Fluorescent Protein (rGFP) from the E. coli strain‚ BL21(DE3) by beginning with its purification via a Ni2+-agarose affinity chromatography column. The His6 tag of the rGFP bound to the Ni2+-agarose column and washes and elutions were obtained
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2. Steps for Southern Blotting Digest the DNA with an appropriate restriction enzyme. Run the digest on an agarose gel. Denature the DNA‚ which would separate double-stranded DNA into single-stranded DNA. Transfer the denatured DNA to the membrane. Probe the membrane. Visualize your radioactively labeled target sequence. If you used a radiolabeled 32P probe‚ then you would visualize by autoradiograph. 3. a) She placed the nucleotide as she did because the EcoR I cuts in one
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Figure 2 also indicated the success of the PCR amplification since we were able to construct the gel map found in Table 1. In Table 1‚ lanes 1‚ 3‚ 6‚ 9‚12 and 14 were expected not to show any DNA since they were not loaded. We used different primers for the positive and negative controls for the sole of validity and accuracy of the 100bp ladder when
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together by weak hydrogen bonds. A DNA fingerprint is manufactured by first extracting the DNA from the cells and multiplying the DNA by performing the polymerase chain reaction (PCR). The DNA is then analysed by use of restriction enzymes and electrophoresis. The DNA is firstly multiplied by the polymerase chain reaction (PCR)‚ which is artificial DNA replication. DNA is extracted from the cells‚ which were possibly found at the crime scene. This DNA is added to a test-tube simultaneously with
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