Gel electrophoresis. See

The sample is loaded at a flow-rate of 1 ml/min onto the FPLC column equilibrated with the same MOPS buffer used to resuspend the RNA pellets. The free nucleotides are completely removed with a 5-ml wash with 350 mM NaCl and the RNA is eluted with a 20-ml (350—750 mM NaCl) linear gradient and analyzed by PAGE/urea gel electrophoresis (see later). Up to 2 mg of RNA can be loaded onto and eluted from a 1-ml (of resin) mono Q column without loss of resolution. The homogeneity of RNA in the fractions collected, as seen by gel electrophoresis, should be >90%. The appropriate fractions are pooled and the RNA collected by ethanol precipitation. The RNA pellet is washed twice with 70% ethanol, air-dried, and finally redissolved in DEPC-treated H20. The total recovery after the entire procedure of purification is = 90%. This protocol yields = 800 pmoles of purified 002 mRNA/pmole template DNA. 

One microliter of the plasmids thus prepared is treated with Sjjfi/Pmel and analyzed by agarose gel electrophoresis see Note 7). Clones with an appropriate insert size are applied to sequencing analysis. Sequencing samples are prepared from the purified plasmids using an illustra TempliPhi DNA Amplification Kit, because the yields of plasmid in a 96-well format preparation are sometimes insufficient for sequencing. 

Several of the synthetic detergents used for dissolving membranes and solubilizing integral membrane proteins. Triton X-100 and octylglucoside are nonionic detergents cetyltrimethylammonium bromide and sodium dodecylsulfate (SDS) are ionic. SDS is also an effective denaturant of proteins and is used in polyacrylamide-gel electrophoresis (see chapter 6). 

Effect of Sialic Acid on SDS Polyacrylamide Gel Electrophoresis Suppose you have four forms of a protein, all with identical amino acid sequence but containing zero, one, two, or three oligosaccharide chains, each ending in a single sialic acid residue. Draw the gel pattern you would expect when a mixture of these four glycoproteins is subjected to SDS polyacrylamide gel electrophoresis (see Fig. 3-18) and stained for protein. Identify any bands in your drawing. 

Rapp with Burtin (R1) found 5 carboxylesterases in the gastric human mucosa on agar gel electrophoresis (see Fig. 11) using special substrates. These carboxyl esterases were decreased and differently distributed in the gastric mucosa of patients with gastric cancer. 

The PCR product is purified on a Seakem LE agarose gel electrophoresis (see Subheading 3.3.5.), The concentration of the gel is 1% instead of 0.5%. The desired PCR fragment is excised from the gel and extracted using the QIAquick Gel Extraction Kit (Westburg) ... 

Restrict the purified plasmid with the appropriate restriction enzyme and analyze the digest by agarose gel electrophoresis (see Note 23). 

Sodium 5-methoxysalicylate 1465 Sodium caprate 1376 Sodium deoxycholate 1465 Sodium dodecyl sulfate-polyacrylamide gel electrophoresis, see SDS-PAGE Sodium glycocholate 1465 Sodium glycodeoxycholate 1376 Sodium salicylate 1376, 1466 Sodium taurodihydrofusidate 1376 Softenon 1675 Solanum lycopersicum 970 Solanum tuberosum 485 Solclot 462 Solid tumor 1174... 

Isoelectric focusing in polyacrylamide gel today appears to be nearly the most popular version of this technique [142-145]. The essential components for the gel preparation are identical with those used with conventional polyacrylamide gel electrophoresis (see pg. 428). The only difference is that carrier ampholytes (2% w/w of the total gel volume) are incorporated into the gel solution before polymerization. The protein sample (free of salts) can be mixed with the sample gel solution or it can be loaded in the conventional way in a sucrose solution. In the latter case it is covered with a protective layer of ampholyte. 

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