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Agarose electrophoresis and

Two isolation procedures based on methods in category 2 are described in this experiment, a large-scale and a microscale method. Each procedure yields plasmid DNA that is sufficiently pure for size analysis by agarose electrophoresis and for digestion by restriction enzymes as described in Experiment 15. [Pg.420]

Soto CM, Blum AS, Wilson CD, Lazordk J, Kim M, Gnade B, Ratna BR (2004) Separation and recovery of intact gold-virus complex by agarose electrophoresis and electroelution application to the purification of cowpea mosaic virus and colloidal gold complex. Electrophoresis 25 2901-2906... [Pg.98]

Figure 2 In ribosome display, mRNA (A) extracted from a cell is converted into a cDNA library (B) is transcribed back into mRNA with no stop codons. Prokaryotic or eukaryotic proteosomes are added and the ribosome then travels down the mRNA (C) translating until it reaches the end of the mRNA molecule (D), where the ribosome halts. With no stop codon, the release factor proteins cannot bind and so the protein, ribosome, and mRNA are physically associated and can be stabilized by high Mg2+ and low temperatures. This complex could then be bound directly to an immobilized natural product (E), the nonbinding library members washed away and the bound members eluted with EDTA (F), which destabilizes the ribosomal complexes by removing Mg2+. The purified sublibrary is converted into cDNA by reverse transcription (RT-PCR) and amplified by regular PCR (B). The/n vitro transcription and translation can be repeated for another round of selection or the cDNA can be analyzed by agarose electrophoresis and/or sequencing. Figure 2 In ribosome display, mRNA (A) extracted from a cell is converted into a cDNA library (B) is transcribed back into mRNA with no stop codons. Prokaryotic or eukaryotic proteosomes are added and the ribosome then travels down the mRNA (C) translating until it reaches the end of the mRNA molecule (D), where the ribosome halts. With no stop codon, the release factor proteins cannot bind and so the protein, ribosome, and mRNA are physically associated and can be stabilized by high Mg2+ and low temperatures. This complex could then be bound directly to an immobilized natural product (E), the nonbinding library members washed away and the bound members eluted with EDTA (F), which destabilizes the ribosomal complexes by removing Mg2+. The purified sublibrary is converted into cDNA by reverse transcription (RT-PCR) and amplified by regular PCR (B). The/n vitro transcription and translation can be repeated for another round of selection or the cDNA can be analyzed by agarose electrophoresis and/or sequencing.
The use of agarose as an electrophoretic method is widespread (32—35). An example of its use is in the evaluation and typing of DNA both in forensics (see Forensic chemistry) and to study heritable diseases (36). Agarose electrophoresis is combined with other analytical tools such as Southern blotting, polymerase chain reaction, and fluorescence. The advantages of agarose electrophoresis are that it requires no additives or cross-linkers for polymerization, it is not hazardous, low concentration gels are relatively sturdy, it is inexpensive, and it can be combined with many other analytical methods. [Pg.182]

The used variants of Cg-AR application were adding to the wells of the gel to DNA, directly bringing into the agarose gel and the addition to electrophoretic buffer. The use of the latest way demonstrated its greatest efficiency by saving up to 1.63 times more DNA preparations if compared with the standard method of electrophoresis, while other ways showed 15.45% increase when Cg-AR was introduced into an agarose gel and 1.63%- when added to the DNA preparations. [Pg.192]

Weiss, GH Garner, M Yarmola, E Bocek, P Chrambach, A, A Comparison of Resolution of DNA Fragments Between Agarose Gel and Capillary Zone Electrophoresis in Agarose Solutions, Electrophoresis 16, 1345, 1995. [Pg.623]

An alternative to nucleic acid isolation by electrophoresis followed by electrophoretic elution is HPLC. There are several advantages in the use of HPLC. It is simple and easy to use and has a high degree of resolution, it is faster than electrophoresis and contamination of DNA fragments by impurities from the agarose gel is avoided. [Pg.454]

Electroimmunoassay (rocket electrophoresis) and radial immunodiffusion (A5) lack sensitivity at low Lp(a) concentrations, and the response is influenced by the size of the apo(a) isoforms (A5, K28). Differences in migration velocity in the agarose gel lead to an underestimation of the samples with large apo(a) isoforms and to an overestimation of samples with small apo(a) isoforms. Moreover, the detection limit lies around 0.07-0.08 g/liter Lp(a), so that this method is better suited for screening and detection of individuals with elevated Lp(a) levels than for the exact measurement of the plasma Lp(a) concentration. [Pg.107]

CNTs can conjugate with nucleic acids via non-covalent bond. ssDNA, short double-stranded DNA and total RNA molecules can attach to the surface of CNTs and can disperse CNTs in aqueous environment. The poly(30T) has the highest dispersion efficiency (Zheng et al., 2003). For example, 1 mg DNA molecules mix with lmg CNTs in 1ml water, yield at most 4mg/ml CNT solution. DNA-CNT complexes can be purified or isolated by electronic properties such as agarose gel electrophoresis and centrifuge method (Cui et al., 2004a Karajanagi et al., 2004). [Pg.183]

Serum is separated by agarose gel electrophoresis and the gel is stained for lipids [45, 74]. Agarose electrophoresis separates four major lipoproteins (i.e., chylomicrons,... [Pg.506]

Clearly a number of differences could explain these divergent results. Boiling could cause loss of antigenicity but retention of activity in stimulating the formation of O . Alternately, the Cab might be attached to agarose beads and to erythrocytes in different ways so that different parts of the molecule were exposed to the PMNs. The assays used to detect contamination of Cab by IgG may also differ in sensitivity (immunofluorescence vs. gel electrophoresis of isolated Ca). [Pg.41]

The resulting DNA was separated by electrophoresis on an agarose gel, and the fluorescent bands on the gel were located. The band pattern resulting from nucleotide mixture 1 is shown below. Assuming that all mixtures were run on the same gel, what did the remaining lanes of the gel look like ... [Pg.304]

Extrachromosomal DNA molecules called plasmids are harbored in some strains of E. coli. The normal copy number of the plasmids is small, between 2 and 10 however, if these strains of E. coli are grown in the presence of chloramphenicol, up to 3000 copies may be replicated per cell. Plasmid DNA has been demonstrated to be a useful vehicle in molecular cloning. This experiment describes a method for the growth of E. coli and amplification of the ColEl plasmids. The plasmids will be isolated from E. coli cells by one of two methods, a large-scale boiling method or a microscale alkaline lysis method. The DNA plasmids will be measured for molecular size by agarose electrophoresis. [Pg.415]


See other pages where Agarose electrophoresis and is mentioned: [Pg.453]    [Pg.31]    [Pg.1422]    [Pg.1423]    [Pg.197]    [Pg.282]    [Pg.453]    [Pg.31]    [Pg.1422]    [Pg.1423]    [Pg.197]    [Pg.282]    [Pg.440]    [Pg.182]    [Pg.182]    [Pg.445]    [Pg.554]    [Pg.585]    [Pg.604]    [Pg.706]    [Pg.10]    [Pg.124]    [Pg.203]    [Pg.111]    [Pg.463]    [Pg.581]    [Pg.377]    [Pg.381]    [Pg.139]    [Pg.142]    [Pg.19]    [Pg.484]    [Pg.30]    [Pg.237]    [Pg.33]    [Pg.98]    [Pg.506]    [Pg.533]    [Pg.145]    [Pg.109]    [Pg.123]    [Pg.126]   


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