LigationOptimization
The following protocol can be used to optimize ligation conditions for difficult to clone (e.g. very large) fragments. The principle is to independently characterize the ligation kinetics of the vector and insert DNA fragments and then to combine them in optimal ratios. The final ligation is also performed in two stages to optimize the proportion of vector:insert heterodimers followed by a shift to low concentration ......阅读全文
Ligation－Optimization
The following protocol can be used to optimize ligation conditions for difficult to clone (e.g. very large) fragments. The principle is to independent
Linker－Ligation
Linker Ligation (with T4 DNA Ligase)In a microcentrifuge tube prepare a solution of blunt ended, dephosphorylated DNA (100-500ng) in TE buffer (5-7µl)
WHITE－PAPER:－Virotherapy－Process－Optimization（一）
WHITE PAPER: Virotherapy Process Optimization采用一系列方法将病毒工程开发用于疾病治疗已经成为一种新型的应用，该应用广泛的定义为病毒疗法，包括用于基因治疗的病毒载体(Figure 1)和具有溶瘤细胞功能的病毒(Figure 2)。虽然这些产品大部分
WHITE－PAPER:－Virotherapy－Process－Optimization（二）
Figure 3. Virus Counter 3100 (top) and Autosampler (bottom)Proof of Principle病毒计数仪已经被广泛应用于多种病毒的计数，包括基因传递载体和溶瘤功能的病毒。这里我们列出一些检测的数据，证实病毒计数仪快速的病毒定量在病毒疗法领域
实验室自动化与筛选协会2013亚洲会展展商技术论坛
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DNA克隆
DNA克隆（主要内容如下）· General Procedure· PCR Cloning· Subcloning· ET Cloning· Vector Preparation· Ligation Re
T载体的制作和应用
Also see DNA Cloning§ Making TA Vector (Crawford Lab)T-vectors are linear-blunt-ended plasmids with a few dT's added on by Taq polymerase.
分子克隆蛋白表达实验指南（四）
7. PCR产物与TA载体连接 pGEM-T vector is T-tailed at the insert site. To improve the ligation efficiency, it is recommended PCR product be A-tailed. +A s
基于epMotion－5075t系统与KPPA－HyperPlus试剂盒的全自动测序...2
Results and DiscussionThe post-ligation qPCR results were used to calculate the percentage of starting material that was successfully adapter ligate
UV－CrossLinking－an...－（二）
实验步骤1. UV cross-linking of tissue culture cells 1) Remove media and add ice-cold PBS to cells (e.g. use cells grown in a 10 cm plate for three ex
DNA测序
DNA测序（主要内容如下）· Sequencing Gel Preparation· Preparation of Templates · DNA Sequencing by the Dideoxy Method· DNA Sequen
利用Mastercycler－X50系列PCR独有的2D梯度实现高效的PCR条件...
利用Mastercycler X50系列PCR独有的2D梯度实现高效的PCR条件优化Ultimate PCR Optimization with Eppendorf Mastercycler® X50 2D-gradientArora Phang, Tim Schommartz,Eppendorf
DNA转化实验指导4
2B. Transformation 1. Preparation of electrocompetent DH5a cells: autoclave 4 baffled 1 liter flasks containing 500 mL LB. Remove a 1 mL aliquo
重组DNA的分离、克隆与测序实验手册5
C. Random fragment end-repair, size selection, and phosphorylationSince both sonicated and nebulized DNA fragments usually contain single-stranded end
DNA的酶学操作
DNA的酶学操作DNA Modifying Enzymes (Michael Blaber)Introduction to bacterial restriction/modification system. It provides very useful background knowledge
DNA转化实验指导1
CONTENT Transformation-Competent E. coli preparation Inoue "ultra-competent" methodRubidium chloride methodCosmid packaging protocol DNA Ligation an
如何利用远红外荧光对细胞内活性氧进行检测
点击次数：10296 发布日期：2018-11-14 来源：本站 仅供参考，谢绝转载，否则责任自负 引言 活性氧 (Reactive oxygen species，ROS) 是 一类由细胞有氧代谢产生的具有化学反应 活性的一类含氧分子，参与细胞信号转 导、免疫防御
含有目的基因真核表达－pEGFPC1载体的构建实验_克隆法
实验方法原理在设计引物时,一对引物两端分别加了两个酶切位点,这两个酶切位点与pEGFP-C1 载体多克隆位点中的酶切位点相吻合,且位置和方向都合适。这样, 目的基因片段和pEGFP-C1 载体经双酶切后,由于酶切位点一致,在T4 DNA 连接酶的作用下就可以连接。实验材料DNA 片段试剂、试剂盒pE
如何利用远红外荧光对细胞内活性氧进行检测
活性氧 (Reactive oxygen species，ROS) 是 一类由细胞有氧代谢产生的具有化学反应 活性的一类含氧分子，参与细胞信号转 导、免疫防御以及维持体内动态平衡等各 项生理活动。然而当细胞处于环境压力 时，胞内的 ROS 含量水平就会急剧上升造 成核酸、蛋白和脂类物质的氧化
如何利用远红外荧光对细胞内活性氧进行检测？
引言活性氧 (Reactive oxygen species，ROS) 是 一类由细胞有氧代谢产生的具有化学反应 活性的一类含氧分子，参与细胞信号转 导、免疫防御以及维持体内动态平衡等各 项生理活动。然而当细胞处于环境压力 时，胞内的 ROS 含量水平就会急剧上升造 成核酸、蛋白和脂类物质的
如何利用远红外荧光对细胞内活性氧进行检测？
活性氧 (Reactive oxygen species，ROS) 是一类由细胞有氧代谢产生的具有化学反应活性的一类含氧分子，参与细胞信号转导、免疫防御以及维持体内动态平衡等各项生理活动。然而当细胞处于环境压力时，胞内的ROS 含量水平就会急剧上升造成核酸、蛋白和脂类物质的氧化损伤[1]，
TOP10－chemically－competent－cells
OverviewThis protocol is a variant of the Hanahan protocol [1] using CCMB80 buffer for DH10B, TOP10 and MachI strains. It builds on Example 2 of the B
2024年4月中国学者被撤回176篇文章－医院是撤稿高发区
在2024年4月，诚信科研编辑部共报道了239篇撤稿文章，其中攘括所有的中国学者撤稿文章及极具影响力的国外单位撤稿文章，包含Lancet，Science，Nature Medicine，Nature Communications，EBioMedicine，Annals of the Rheuma
概述食管静脉曲张的治疗原则
内镜下食管静脉曲张套扎术（endoscopic esophageal varix ligation，EVL）是20世纪80年代发展起来的治疗EVB的重要手段，它将橡胶圈安装于内镜前端，将曲张静脉负压吸入透明帽后用橡胶圈套扎，通过机械作用使曲张静脉血流中断，血栓形成，最后发生坏死溃疡，愈合遗留瘢痕
UV－CrossLinking－an...－（一）
实验概要Interest in RNA-protein interactions is booming as we begin to appreciate the role of RNA, not just in well-established processes such as trans
DNA的诱变和甲基化
· In Vitro Mutagenesis Using Altered Sites (Bowtell Lab) In vitro Mutagenesis with dut ung single stranded DNA (Hahn Lab)· Site-direct
Construction－of－BAC－Libraries：Construction－of－a－BAC－library
Once high molecular weight (HMW) DNA has been prepared it must somehow be fragmented and DNA in the desired size range isolated. In general, as the de
基于epMotion－5075t系统与KPPA－HyperPlus试剂盒的全自动测序..1
基于epMotion 5075t系统与KPPA HyperPlus试剂盒的全自动测序前文库制备方案Automated KAPA HyperPlus DNA Library Preparation for Illumina® Sequencing on the Eppendorf epMotion®
CTCF:－First－Multivalent－Nuclear－Factor
CTCF is central to signaling pathways in immature B cells elicited by cross-linking the Ig BCR and stimulation with TGF?. Both stimuli result in induc
Transient－Transfection－of－Cos1－Cells
Transient Transfection of Cos-1 CellsNote: This protocol has been optimized for Cos-1 cells. Successful transfection of each cell type requires optimi