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1.
Synthesis of nucleic acid probes on membrane supports: a procedure for the removal of unincorporated precursors 总被引:1,自引:0,他引:1
S P Bhat 《Analytical biochemistry》1990,184(1):59-62
We have used DNA bound to small pieces of nylon membrane for the synthesis of radioactive probes. The DNA to be used for generating the probe(s) is first bound to nylon membranes and then introduced into the reaction mix. The labeling reaction takes place on the membrane and therefore allows easy removal of unincorporated precursors by simple washing for 1-2 min. The clean labeled probe is eluted from the membrane in formamide or in water and is ready for use. This DNA-membrane can be stored for reuse. Synthesis of probes on a solid support such as nylon membrane thus circumvents problems associated with chromatographic manipulations needed for the separation of labeled DNA from unicorporated precursors. Probes synthesized in this manner are as efficient in detecting nucleic acid sequences as those synthesized in solution. 相似文献
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Anwesha Dutta Premalatha Shetty Smitha Bhat Yeshaswini Ramachandra Shrinidhi Hegde 《Journal of biomolecular techniques》2012,23(4):128-135
A solvent system that extracts a maximum number of metabolites belonging to diverse chemical classes from complex biofluids, such as plasma, may offer useful inputs to understand the metabolic and physiological state of an individual. The present study compared seven solvent systems for extraction of metabolites from plasma. The extracts were analyzed by mass spectrometry (MS) and MS/MS (MS2) using a quadrupole time-of-flight liquid chromatography/MS system in positive and negative modes of ionization. Metabolites with molecular mass below 400 were identified using Human Metabolome Database MS2 and MS search interfaces. The acetone/isopropanol (2:1) system yielded promising results in positive ionization mode, as the maximum number of MS and MS2 features was detected in the extract. It was found to be superior in extraction of various classes of metabolites, especially organic acids, nucleosides and nucleoside derivatives, and heterocyclic molecules. Glycerophosphocholines in the mass range of 400–700 were found to be efficiently extracted by the methanol/chloroform/water (8:1:1) system. In negative mode as well, the maximum number of MS2 features was detected in methanol/chloroform/water and acetone/isopropanol extracts. The fingerprints of molecular features obtained in the negative and positive modes differed from each other to a significant extent. 相似文献
4.
Audesh Bhat Anil Koul Ekta Rai Swarkar Sharma M. K. Dhar R. N. K. Bamezai 《Human genetics》2008,123(1):115
5.
Haploid plants were regenerated in vitro from unpollinated ovules of niger (Guizotia abyssinica (L. f.) (Cass.) on Murashige and Skoog nutrient medium (MS) supplemented with 10 μM naphthaleneacetic acid or 10 μM NAA +
1.5 μM kinetin and 30 g/l sucrose. Gamborg (B5) medium was the best for plant regeneration (in comparison with MS, Nitsch
and Nitsch (NN), and Chu (N6) media) from cultured ovules, and 6.66 and 7.33 ovules of JNC-6 and Ootacamund cultivars were
involved in direct plant regeneration on this medium. Matured ovules (ovules collected one day before anthesis or on the day
of anthesis) only responded to cultural regimes and involved in direct plantlet development. Cytological preparation of root
tips and chloroplast counts in the guard cells of leaf stomata of regenerated plants confirmed their haploid nature.
This text was submitted by the authors in English. 相似文献
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Isolation and screening of potential actinobacteria for rapid composting of rice straw 总被引:2,自引:0,他引:2
Rice straw is produced as a by-product from rice cultivation, which is composed largely of lignocellulosic materials amenable
to general biodegradation. Lignocellulolytic actinobacteria can be used as a potential agent for rapid composting of bulky
rice straw. Twenty-five actinobacteria isolates were isolated from various in situ and in vitro rice straw compost sources.
Isolates A2, A4, A7, A9 and A24 were selected through enzymatic degradation of starch, cellulose and lignin followed by the
screening for their adaptability on rice straw powder amended media. The best adapted isolate (A7) was identified as Micromonospora
carbonacea. It was able to degrade cellulose, hemicelluloses and carbon significantly (P ≤ 0.05) over the control. C/N ratio was reduced to 18.1 from an initial value of 29.3 in 6 weeks of composting thus having
the potential to be used in large scale composting of rice straw. 相似文献
8.
Hasidah Mohd. Sidek Cynthia Nyquist-Battie Garret Vanderkooi 《Biochimica et Biophysica Acta (BBA)/General Subjects》1984,801(1):26-31
The effects of tertiary amine local anesthetics (procaine, lidocaine, tetracaine and dibucaine) and chlorpromazine were investigated for three enzyme activities associated with rat brain synaptosomal membranes, i.e., (Na+ + K+)-ATPase (ouabain-sensitive), Mg2+-ATPase (ouabain-insensitive) and acetylcholinesterase. Approximately the same concentrations of each agent gave 50% inhibition of both ATPase, for example 7.9 and 10 mM tetracaine for Mg2+-ATPase and (Na+ + K+)-ATPase, respectively; these concentrations are 10-fold higher than required for inhibition of mitochondrial F1-ATPase. The relative inhibitory potency of the several agents was proportional to their octanol/water partition coefficients. Acetylcholinesterase was inhibited by all agents tested, but the ester anesthetics (procaine and tetracaine) were considerably more potent than the others after correction for partition coefficient differences. For tetracaine, 0.18 mM gave 50% inhibition and showed competitive inhibition on a Lineweaver-Burk plot, but for dibucaine a mixed type of inhibition was observed, and 0.63 mM was required for 50% inhibition. Tetracaine evidently binds at the active site, and dibucaine at the peripheral or modulator site, on this enzyme. 相似文献
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