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TES buffer

Fig. 3. Gallery of representative nucleosomes reconstituted in the absence (a) or presence of GH5 (b) or H5 (c), and visualized by scanning transmission electron microscopy, (a) and (b) 256 bp 5S rDNA fragment [65]. (c) 357 bp fragment from the 5S series (see text). Samples were diluted in TE buffer supplemented with 50 mM NaCl and 5 mM MgCl2 before adsorption to the grids. Note the nucleosome different positions relative to the DNA ends. Bars 25 nm and 75 bp. (Adapted from Fig. 7,9, and 10 in Ref [34].) Schemes of the corresponding DNA conformations are shown. Fig. 3. Gallery of representative nucleosomes reconstituted in the absence (a) or presence of GH5 (b) or H5 (c), and visualized by scanning transmission electron microscopy, (a) and (b) 256 bp 5S rDNA fragment [65]. (c) 357 bp fragment from the 5S series (see text). Samples were diluted in TE buffer supplemented with 50 mM NaCl and 5 mM MgCl2 before adsorption to the grids. Note the nucleosome different positions relative to the DNA ends. Bars 25 nm and 75 bp. (Adapted from Fig. 7,9, and 10 in Ref [34].) Schemes of the corresponding DNA conformations are shown.
Fig. 4. Nucleosome relaxation, and influence of histone N-terminal tails. Example of nucleosomes on 356 bp ALk= —2.9 topoisomer from the pBR DNA minicircle series [28]. (a) Mononucleosomes (Mo) were reconstituted with control (Control) or acetylated (Acetyl) histones, incubated at 37 °C in Tris buffer [T 50 mM Tris-HCl (pH 7.5), 0.1 mM EDTA, 50 mM KCl, 5 mM MgC, and 0.5 mM dithiothreitol] or phosphate buffer [P same as Tris buffer with 50 mM potassium phosphate (pH 7.5) instead of 50 mM Tris-HCl] in the absence (Topo I —) or presence (Topo I +) of topoisomerase I, and electrophoresed in a native polyacrylamide gel at room temperature. Note the splitting of nucleosome relaxation products in two bands. TE starting chromatin in TE buffer, (b) Gel slices (brackets) were cut out, and eluted DNAs were electrophoresed in a chloroquine-containing native polyacrylamide gel, together with control naked topoisomers (C1-C4). Lanes were numbered as in the (a) gel. Autoradiograms are shown, (c) Radioactivity profiles of lanes 2 and 5 in the (b) gel. Topoisomers are indicated by their ALk values. (Adapted from Fig. 2 in Ref. [28].)... Fig. 4. Nucleosome relaxation, and influence of histone N-terminal tails. Example of nucleosomes on 356 bp ALk= —2.9 topoisomer from the pBR DNA minicircle series [28]. (a) Mononucleosomes (Mo) were reconstituted with control (Control) or acetylated (Acetyl) histones, incubated at 37 °C in Tris buffer [T 50 mM Tris-HCl (pH 7.5), 0.1 mM EDTA, 50 mM KCl, 5 mM MgC, and 0.5 mM dithiothreitol] or phosphate buffer [P same as Tris buffer with 50 mM potassium phosphate (pH 7.5) instead of 50 mM Tris-HCl] in the absence (Topo I —) or presence (Topo I +) of topoisomerase I, and electrophoresed in a native polyacrylamide gel at room temperature. Note the splitting of nucleosome relaxation products in two bands. TE starting chromatin in TE buffer, (b) Gel slices (brackets) were cut out, and eluted DNAs were electrophoresed in a chloroquine-containing native polyacrylamide gel, together with control naked topoisomers (C1-C4). Lanes were numbered as in the (a) gel. Autoradiograms are shown, (c) Radioactivity profiles of lanes 2 and 5 in the (b) gel. Topoisomers are indicated by their ALk values. (Adapted from Fig. 2 in Ref. [28].)...
Dilute IMMEDIATELY with 40 p, TE Buffer (10 mM Tris, pH 8.0,1 mM EDTA) and either transform IMMEDIATELY or freeze the reaction until you are ready to transform. coli with a transformation efficiency in excess of 1 x 10 are recommended and if the vector has been modified for blue-white screening ensure that an appropriate E. coli host strain is used 5 pi of the diluted reaction should give tens to hundreds of colonies per well of a 24-well plate. [Pg.28]

Tris-EDTA (TE) buffer 10 mM TRIZJMA hydrochloride, 1 mM EDTA. [Pg.24]

B 1 pg/ml ethidium bromide in TE buffer by dilution of a stock solution of 1 mg/ml EtBr in Soln. A. (The other mentioned dyes are used the same way.)... [Pg.16]

TBS maybe made as tenfold stock and stored frozen in aliquots. Sometimes you will find the following composition for TBS 12.11 g Tris (0.1 Mole), 2.05 g NaCl (35 mMoles), 0.75 g glycine (10 mMoles), 0.2 gNaNs or Thimerosalper 1000 ml,pH7.8-8.4. TE-Buffer... [Pg.204]

A tenfold stock of TE buffer is possible. Check pH after dilution to working concentration, because the pK of Tris has a relatively large temperature and concentration dependency. [Pg.204]

For 96 samples, prepare 100 samples premix. Assemble the following recipe Donor vector (pDONR221 150 ng/pl) 50 pi, BP reaction buffer (5x) 100 pi, TE buffer 150 pi, BP Clonase enzyme mix 100 pi. [Pg.20]

The plasmid preparation using the Wizard SV 96 Plasmid DNA Purification System follows the protocol from Promega, except that TE buffer instead of H O is used for the final plasmid elution buffer. This alteration is applied to all the plasmid preparation processes in this chapter. [Pg.36]

After the purification, measure the absorbance at 260 nm of the purified mRNA solution using a spectrophotometer. Dilute 2 pL of the purified mRNA solution into 500 pL of TE buffer, and then measure the absorbance. Use TE as the blank. The mRNA concentration is determined by the following equation mRNA concentration (mg/mL) = value X 0.04 x 250. [Pg.104]

N-Terminal nucleophile hydrolases autoactivation of 621 Termites, protozoa in 19 Tertiary structure of a protein 59 TES buffer 99... [Pg.934]

TE buffer Dissolve 10 mM Tris-HCl and 1 mMEDTA (pH 8.0) in sterile distilled water Do not autoclave. [Pg.407]

Remove the reaction mixture from beneath the mineral oil, phenol/chloroform extract, chloroform extract, and precipitate the DNA by adding 25 pL 7 5M ammonium acetate and 150 pL ethanol. Precipitate at room temperature for 10 min. Centrifuge for 10 mm at 12,000 rpm in a microcentrifuge, wash the DNA pellet with 100 pL 70% ethanol, dry, and resuspend in 20 pL TE buffer Digest the DNA fragment with YhoI according to the supplier s instructions. [Pg.433]

Precipitate one-half of the digested vector DNA with ethanol Add 0 5 vol of 7.5M ammonium acetate and 2 vol of 100% ethanol. Incubate at-20°C for 30 min, then spin at full speed in a microcentrifuge for 10 min, wash the pellet with 70% ethanol, dry, and resuspend in 10 pL of TE buffer... [Pg.433]


See other pages where TES buffer is mentioned: [Pg.504]    [Pg.168]    [Pg.206]    [Pg.207]    [Pg.208]    [Pg.210]    [Pg.56]    [Pg.294]    [Pg.581]    [Pg.590]    [Pg.444]    [Pg.457]    [Pg.50]    [Pg.27]    [Pg.314]    [Pg.16]    [Pg.16]    [Pg.14]    [Pg.18]    [Pg.18]    [Pg.21]    [Pg.21]    [Pg.27]    [Pg.100]    [Pg.170]    [Pg.329]    [Pg.329]    [Pg.458]    [Pg.458]    [Pg.380]    [Pg.407]    [Pg.410]    [Pg.410]    [Pg.431]   
See also in sourсe #XX -- [ Pg.15 , Pg.204 ]

See also in sourсe #XX -- [ Pg.28 ]

See also in sourсe #XX -- [ Pg.28 ]




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