{{'Search' | translate}}
 

Pwo DNA Polymerase

Pwo DNA聚合酶

公司名称: Roche Diagnostics
产品编号: 11644955001
Bio-protocol()
Company-protocol()
Other protocol()

Ultradeep Pyrosequencing of Hepatitis C Virus to Define Evolutionary Phenotypes
Author:
Date:
2017-05-20
[Abstract]  Analysis of hypervariable regions (HVR) using pyrosequencing techniques is hampered by the ability of error correction algorithms to account for the heterogeneity of the variants present. Analysis of between-sample fluctuations to virome sub-populations, and detection of low frequency variants, are unreliable through the application of arbitrary frequency cut offs. Cumulatively this leads to an underestimation of genetic diversity. In the following technique we describe the analysis of Hepatitis C virus (HCV) HVR1 which includes the E1/E2 glycoprotein gene junction. This procedure describes ... [摘要]  使用焦磷酸测序技术的高变区(HVR)分析受到纠错算法解释存在的变异异质性的能力的阻碍。通过应用任意频率切断,对样本间波动与色情子群体的分析以及低频变体的检测是不可靠的。累积地导致遗传多样性的低估。在以下技术中,我们描述了包含E1 / E2糖蛋白基因连接的丙型肝炎病毒(HCV)HVR1的分析。该程序描述了HCV在治疗初始环境中的演变,从10年来收集的10个样品中,使用在Roche GS FLX钛平台上进行的超深度焦磷酸测序(UDPS)(2014年,Palmer等人) 。使用血清样品的初步克隆分析来通知允许达到更大序列深度的下游误差校正算法。已经针对HCV基因型1,2,3和4测试了该区域的PCR扩增。

背景 衍生自病毒扩增子的UDPS数据集的分析经常依赖于未针对扩增子分析进行优化的软件工具,假设随机并入测序突变,并且集中在找到真实序列而不是假变异体。存在于RNA病毒基因组中的高变区存在这些困难。许多利用UDPS的研究通过对数据应用任意的频率切断来寻求解决这些问题,从而导致小的变体的丢失。在这里,暂时匹配的克隆数据集以及旨在克服所概述的问题的纠错方法,有助于保留有价值的序列信息。

Generation of Non-typeable Haemophilus influenzae Directed Gene Deletion Mutants
Author:
Date:
2014-03-05
[Abstract]  Directed deletion mutants in non-typeable Haemophilus influenzae can be made by allelic exchange of the target gene with an artificial DNA construct in which an antibiotic resistance cassette is placed between two ~1,000 bp DNA sequences that are identical to the 5' and 3' flanking regions of the target gene. The artificial DNA construct that is required for this mutagenesis is synthesized by the so-called Megaprimer PCR method (Figure 1).
[摘要]  在非典型流感嗜血杆菌中的定向缺失突变体可以通过靶基因与人工DNA构建体的等位基因交换来制备,其中抗生素抗性盒置于两个〜1000bp的DNA序列之间, 靶基因的5'和3'侧翼区。 该诱变所需的人工DNA构建体通过所谓的Megaprimer PCR方法合成(图1)。

产品评论