{{'Search' | translate}}
 

QIAamp DNA Mini Kit

QIAamp DNA Mini Kit

Company: QIAGEN
Catalog#: 51306
Bio-protocol()
Company-protocol()
Other protocol()

Quantification of HIV RNA and Human Herpesvirus DNA in Seminal Plasma
Author:
Date:
2015-05-05
[Abstract]  Multiple viruses can co-infect the genital tract, modifying the immunologic and virologic milieu and possibly playing a role in viral transmission and pathogenesis. The aim of our studies has been to understand the complex relationships between HIV-1 RNA, and multiple human herpesviruses known to frequently replicate in the genital tract of HIV-infected men (i.e. cytomegalovirus [CMV], Epstein Bar virus [EBV], herpes simplex virus [HSV] types 1 and 2, and human herpesviruses [HHV] 6, 7 and 8) (Gianella et al., 2013a; Gianella et al., 2013b; Gianella et al., 2013c; Gianella et al., 2014). This protocol was designed to collect and process male genital secretion (GS), and to isolate and further quantify HIV RNA and DNA of seven HHV from seminal ... [摘要]  多种病毒可以共感染生殖道,修改免疫学和病毒学环境,并可能在病毒传播和发病中起作用。 我们的研究的目的是了解HIV-1 RNA和已知在HIV感染的男性生殖道中经常复制的多种人疱疹病毒之间的复杂关系(即巨细胞病毒[CMV],Epstein 条形病毒[EBV],单纯疱疹病毒[HSV] 1型和2型,以及人疱疹病毒[HHV] 6,7和8)(Gianella等人,2013a; Gianella等人 。,2013b; Gianella 等人,2013c; Gianella ,2014)。 该方案设计用于收集和处理男性生殖器分泌(GS),并使用定量实时PCR技术从精浆中分离并进一步定量7个HHV的HIV RNA和DNA。

DNA Slot Blot Repair Assay
Author:
Date:
2015-04-20
[Abstract]  Ultraviolet (UV) irradiation induces helix distorting photolesions such as cyclobutane pyrimidine dimers (CPD) and pyrimidine-pyrimidone (6-4) photoproducts (6-4PP) which threaten genomic integrity if unrepaired. In mammals, nucleotide excision repair (NER) is the only pathway that removes UV-induced DNA damages. Here we describe DNA slot blot repair assay for quantitative detection of NER activity using DNA damage specific antibodies such as anti-CPD and anti-6-4PP. Briefly, genomic DNA irradiated with UV was isolated from cells, and the genomic DNA was vacuum-transferred to a nitrocellulose membrane using a Bio-Dot SF microfiltration apparatus (Bio-Rad). A monoclonal antibody that recognizes CPD or 6-4PP was applied to detect the remaining amount of photolesions in the genomic DNA. For ... [摘要]  紫外线(UV)照射诱导螺旋扭曲光致损伤,例如环丁烷嘧啶二聚体(CPD)和嘧啶 - 嘧啶酮(6-4)光产物(6-4PP),如果未修复则威胁基因组完整性。 在哺乳动物中,核苷酸切除修复(NER)是去除UV诱导的DNA损伤的唯一途径。 在这里我们描述DNA狭缝印迹修复测定NER活性使用DNA损伤特异性抗体如抗CPD和抗6-4PP的定量检测。 简言之,从细胞中分离用UV照射的基因组DNA,使用Bio-Dot SF微量过滤装置(Bio-Rad)将基因组DNA真空转移到硝酸纤维素膜上。 应用识别CPD或6-4PP的单克隆抗体来检测基因组DNA中残留的光损伤量。 对于均匀负载的上样控制,可以通过SYBR金染色进一步分析DNA在膜上。

Comments