产品搜索
商品搜索:
购物车
购物车中有 0 件商品 去结算 我的订单
点评详情
发布于:2018-6-11 10:50:19  访问:117 次 回复:0 篇
版主管理 | 推荐 | 删除 | 删除并扣分
Ffer. Soon after electrophoresis, the RNA was visualized by staining with SYBR
The fluorescence was recorded on a spectraMax fluorescence microplate reader (Molecular Device) and the excitation and emission wavelengths were 520 and 560 nm, respectively.Analysis of viral DNA in infected cellsThe MT4 cells (1 ? 105) were with infected HIV-1 corresponding to 20 ng of HIV-1 p24, unless specified otherwise, then harvested after 6 h. The total Doravirine site cellular DNA was extracted making use of the DNeasy blood and tissue kit (Qiagen, Hilden, Germany) as outlined by the manufacturer‘s instruction and analyzed by qPCR applying the following HIV-1- particular oligonucleotides, Forward (5-CAAGTAGTGTGTGCCCGTCTGTT-3), Reverse (5-CTG CTAGAGATTTTTCCACACTGAC-3).Kim et al. Retrovirology (2015) 12:Page 13 ofViral RNA analysisHIV1 reverse transcriptase assayThe total RNA after viral infection was extracted with Trizol reagent (Invitrogen, Carlsbad, CA) as outlined by the manufacturer‘s directions. The viral RNA was analyzed using a northern blot assay. A total of three g of RNA was denatured at 68 in sample buffer containing six.5 formaldehyde, 50 formamide, 1X MOPS, 5 glycerol, and 0.04 bromophenol blue for 15 min and separated on a 0.8 agarose gel containing 2.2 M formaldehyde. This was then transferred to nylon membrane (Roche, Indianapolis, IN). Following transfer, the RNA was fixed towards the membrane by UV cross-linking. For hybridization, a digoxigenin (DIG)-labeled riboprobe system was utilized. The riboprobes which corresponded for the Gag (790?1285 nt) sequences of pNL43GFP have been synthesized by in vitro transcription making use of T7 polymerase in line with the instruction of your Dig Northern Starter Kit (Roche, Indianapolis, PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/26740125 IN), and also the actin probe offered within the kit was made use of. PubMed ID:http://www.ncbi.nlm.nih.gov/pubmed/27689333 The hybridization and detection were performed in accordance with the manufacturer‘s instructions.HIV1 Core uncoating assayThe RT assays were performed using a non-radioactive fluorometric approach [53]. The poly(A) substr.Ffer. Soon after electrophoresis, the RNA was visualized by staining with SYBR?Green I nucleic acid gel-stain answer (Molecular Probes. Carlsbad, CA) The RNA-binding protein was visualized by staining with SYPRO?Ruby EMSA protein gel-stain resolution (Molecular Probes. Carlsbad, CA).cTAR DNA destabilizationFor the SPR assay, the NC protein (GenScript, Piscataway, NJ) was immobilized to a CM5 chip contained inside a 10 mM sodium acetate buffer (pH five.five) remedy. The test compounds had been diluted in 5 DMSO and permitted to flow on chip in PBS operating buffer in the rate of 30 L per min for 120 s. The affinity was measured making use of a Biacore T100 instrument and the data evaluation was processed to acquire all the binding strength and kinetic parameters with all the evaluation plan supplied by the manufacturer (GE Healthcare, USA) [50, 51]. The tryptophan quenching of NC (five M) was performed inside a buffer containing 10 mM sodium phosphate and 10 glycerol with or without the need of A1752. The decrease in fluorescence was measured at excitation and emission wavelengths of 280 and 340 nm, respectively, using a spectraMax Gemini Em fluorescence microplate reader (Molecular Device).The doubly-labeled cTAR DNA oligonucleotides (55 nts) were synthesized at a 0.2 mol scale and purified by the manufacturer (Bioneer Inc., Daejeon, Korea).
共0篇回复 每页10篇 页次:1/1
共0篇回复 每页10篇 页次:1/1
我要回复
回复内容
验 证 码
看不清?更换一张
匿名发表 
点评详情
脚注信息
Copyright (C) 2009-2018 All Rights Reserved. 啄尔雅芝化妆品网上商城 版权所有  
服务时间:周一至周日 08:30 — 20:00  全国订购及服务热线 400 600 6461
联系地址:上海市金山区   邮政编码:310000   QQ:1665638575