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Gel electrophoresis agarose

Usually 1.5% agarose gels are used. More concentrated ones (7-8%) approach, as regards their separation properties, polyacrylamide gels. Separations are done in [Pg.444]

07S M barbital-sodium barbital buffer (pH 8.6) (for plasma proteins) containing 3 mM calcium lactate. The gels are prepared by suspending 1.5 g of agarose in 100 ml of barbital buffer and the suspension is heated until a clean solution is obtained. Then the agarose solution is cooled to 60°C, and 25 ml are poured over a [Pg.444]

The electrophoretic techniques discussed up to this point are useful for analyzing proteins and small fragments of nucleic acids up to 350,000 daltons (500 bp) in molecular size however, the small pore sizes in the gel are not appropriate for analysis of large nucleic acid fragments or intact DNA molecules. The standard method used to characterize RNA and DNA in the range 200 to 50,000 base pairs (50 kilobases) is electrophoresis with agarose as the support medium. [Pg.122]

Agarose, a product extracted from seaweed, is a linear polymer of galac-topyranose derivatives. Gels are prepared by dissolving agarose in warm elec- [Pg.122]

The human genome project is a federal government-sponsored program to sequence all DNA in human chromosomes. [Pg.122]

Nucleic acids can be visualized on the slab gel after separation by soaking in a solution of ethidium bromide, a dye that displays enhanced fluorescence when intercalated between stacked nucleic acid bases. Ethidium bromide may be added directly to the agarose solution before gel formation. This method allows monitoring of nucleic acids during electrophoresis. Irradiation of ethidium bromide-treated gels by UV light results in orange-red bands where nucleic acids are present. [Pg.123]

An apparatus for horizontal slab gel electrophoresis. Courtesy of Hoefer Pharmacia Biotech Inc., San Francisco. [Pg.123]


Eor example, the technique of Southern blotting was developed (68) for use with agarose gel electrophoresis of DNA fragments. Southern blots are designed to detect specific sequences of DNA. After electrophoresis is complete, the DNA is denatured and the single stranded DNA transferred to the specially prepared nitrocellulose paper. The nitrocellulose is then incubated with radioactive RNA or DNA complementary to those DNA sequences of interest. After the nitrocellulose has been sufftciendy incubated with the radioactive complementary DNA, autoradiography is used to identify the fragments of interest. [Pg.184]

Small-angle X-ray scattering (SAXS), circular dichroism (CD), and UV spectroscopy at different temperatures were used to investigate the nature of calf-thymus DNA in aqueous solution, in the presence of [Me Sn] " (n = 1-3) species. The results demonstrate that the [MeSn(IV)] moiety does not influence the structure and conformation of the DNA double helix, and does not degrade DNA, as indicated by agarose gel electrophoresis. Inter alia, the radii of gyration, Rg, of the cross section of native calf-thymus DNA, determined by SAXS in aqueous solution in the presence of [Me Sn] " (n = 1-3) species are constant and independent of the nature and concentration of the [Me Sn] species. [Pg.383]

Grendemann, D., Schumig, E. Protection of DNA during preparative agarose gel electrophoresis against damage induced by ultraviolet light. BioTechniques, Vol.21, No.5, (November 1996), pp. 898-903, ISSN 0736-6205... [Pg.198]

Presented below are four increasingly stringent confirmatory techniques for PCR and a brief discussion of considerations, limitations and advantages of each. These four techniques are agarose gel electrophoresis, restriction analysis. Southern blotting and sequencing. [Pg.664]

Agarose gel electrophoresis can be used to determine whether the PCR amplicon is the expected size. The density of the gel should be chosen to ensure resolution of... [Pg.664]

Calf thymus (CT) DNA was first fragmented by sonication, then purified by a standard procedure. Agarose gel electrophoresis for the sonicated CT DNA revealed a distribution in molecular weight ranging from 30 kDa to 260 kDa which is equivalent in base pairs (bp) from 40 to 400 (1 bp = 660 Da). The DNA fragments were reacted with HEDS in the presence of l-cyclohexyl-3-(2-morpholinoethyl) carbodi-imide metho-p-toluenesulfonate (CMC/jTs). Finally, the reaction mixture was gel-filtered and the macro-molecular fractions, when displayed the characteristic absorption of 260 nm of the nucleic bases, were collected. [Pg.519]

Necrosis by LDH Mitochondrial transmembrane potential by DiOC6 and JC-1 fluorescence Apoptosis by cytochrome c release Annexin V binding DNA fragmentation by agarose gel electrophoresis... [Pg.546]

Experiment 62 Separation of Hemoglobin and Cytochrome C by Horizontal Agarose Gel Electrophoresis... [Pg.483]

Why is agarose gel electrophoresis generally preferable to polyacrylamide gel electrophoresis ... [Pg.486]

Agarose gel electrophoresis can only be used for DNA molecules that are greater than 200 base pairs in size. For molecules smaller than this polyacrylamide gel... [Pg.452]

Fig. 5. Thirty-four out of 35 potential apo(a) isoforms, separated by SDS-agarose gel electrophoresis. The photograph represents a composite of two separate gels with the reference mixture (St) and 17 different samples applied to each gel. Samples were selected to represent each of the observed isoforms. Twenty-nine samples had single-handed patterns and five samples had double-banded patterns. The double banded types and every fifth single-handed phenotype are indicated at the bottom of each gel lane. [With permission of Marcovina et al. (M12).]... Fig. 5. Thirty-four out of 35 potential apo(a) isoforms, separated by SDS-agarose gel electrophoresis. The photograph represents a composite of two separate gels with the reference mixture (St) and 17 different samples applied to each gel. Samples were selected to represent each of the observed isoforms. Twenty-nine samples had single-handed patterns and five samples had double-banded patterns. The double banded types and every fifth single-handed phenotype are indicated at the bottom of each gel lane. [With permission of Marcovina et al. (M12).]...
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]

Separate products by agarose gel electrophoresis Slain with cl Iridium bromide and scan with suitable imaging system c.g. Kodak EDAS 2 0... [Pg.344]


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