Synthetic DNA Assembly Using Golden Gate Cloning and the Hierarchical Modular Cloning Pipeline [0.03%]
使用金门克隆和分级模块克隆流水线进行合成DNA组装
Sylvestre Marillonnet,Ramona Grützner
Sylvestre Marillonnet
Methods that enable the construction of recombinant DNA molecules are essential tools for biological research and biotechnology. Golden Gate cloning is used for assembly of multiple DNA fragments in a defined linear order in a recipient vec...
Amplification Techniques for the Detection of Misfolded Prion Proteins in Experimental and Clinical Samples [0.03%]
实验样品和临床样品中错误折叠的朊蛋白检测的放大技术
Nicholas Haley
Nicholas Haley
This article describes two methods for amplifying prions present in experimental and clinical samples: the protein misfolding cyclic amplification (PMCA) assay and the real-time quaking-induced conversion (RT-QuIC) assay. Protocols for prep...
Multigene Engineering by GoldenBraid Cloning: From Plants to Filamentous Fungi and Beyond [0.03%]
黄金拼接克隆的多基因工程：从植物到丝状真菌及其应用
Marta Vazquez-Vilar,Mónica Gandía,Victor García-Carpintero et al.
Marta Vazquez-Vilar et al.
Many synthetic biologists have adopted methods based on Type IIS restriction enzymes and Golden Gate technology in their cloning procedures, as these enable the combinatorial assembly of modular elements in a very efficient way following st...
Somatic Liver Knockout (SLiK): A Quick and Efficient Way to Generate Liver-Specific Knockout Mice Using Multiplex CRISPR/Cas9 Gene Editing [0.03%]
SLiK肝脏基因敲除技术：一种利用多重CRISPR/Cas9基因编辑快速高效产生肝特异性基因敲除小鼠的新方法
Collin G Johnson,Tong Chen,Nika Furey et al.
Collin G Johnson et al.
Somatic liver knockout (SLiK) is a method developed to rapidly generate a liver-specific knockout of one or several genes. This technique combines the strengths of CRISPR/Cas9 gene editing and hydrodynamic tail-vein injection, a simple in v...
How to Perform miRacles: A Step-by-Step microRNA Detection Protocol Using DNA Nanoswitches [0.03%]
用DNA纳米开关检测microRNA的步步奇迹法
Arun Richard Chandrasekaran,Bijan K Dey,Ken Halvorsen
Arun Richard Chandrasekaran
MicroRNAs are short non-coding RNAs involved in post-transcriptional gene regulation, and are increasingly considered to be biomarkers for numerous biological processes and human diseases. Current techniques used for microRNA detection can ...
Qingqing Yan,Kavitha Sarma
Qingqing Yan
R-loops are abundant, RNA-containing chromatin structures that form in the genomes of both eukaryotes and prokaryotes. Devising methods to identify the precise genomic locations of R-loops is critical to understand how these structures regu...
Use of the CRISPR-Cas9 System in Drosophila Cultured Cells to Introduce Fluorescent Tags into Endogenous Genes [0.03%]
应用CRISPR-Cas9系统在果蝇培养细胞中对内源性基因进行荧光标记
Justin A Bosch,Shannon Knight,Oguz Kanca et al.
Justin A Bosch et al.
The CRISPR-Cas9 system makes it possible to cause double-strand breaks in specific regions, inducing repair. In the presence of a donor construct, repair can involve insertion or 'knock-in' of an exogenous cassette. One common application o...
Rapid, Efficient, and Modular Generation of Adenoviral Vectors via Isothermal Assembly [0.03%]
基于等温组装的快速、高效和模块化的腺病毒载体构建方法
Yong Yang,Yudan Chi,Xinying Tang et al.
Yong Yang et al.
Adenoviral vectors have yielded promising results as carriers for gene transfer and vaccines in basic research and clinical applications. However, most common procedures to construct adenoviral vectors and manipulate adenovirus (Ad) genomes...
Selective Depletion of Abundant RNAs to Enable Transcriptome Analysis of Low-Input and Highly Degraded Human RNA [0.03%]
选择性贫化丰富RNA以实现低输入和高度降解的人类RNA转录组分析
Daniela B Munafó,Bradley W Langhorst,Christine L Chater et al.
Daniela B Munafó et al.
Ribosomal RNAs (rRNAs) are extremely abundant, often constituting 80% to 90% of total RNA. Since rRNA sequences are often not of interest in genomic RNA sequencing experiments, rRNAs can be removed from the sample before the library prepara...
Single-Cell Genome and Transcriptome Sequencing Library Construction Using Combination of MDA and Nextera Library Prep Method [0.03%]
结合MDA与Nextera文库制备法的单细胞基因组及转录组测序文库构建方法
Ioanna Andreou,Christian Korfhage,Nan Fang
Ioanna Andreou
Single-cell analysis gives insights into the heterogeneity of neighboring cells within tissues or within cell populations and is increasing in importance in life science and medicine. Genome and transcriptome sequencing require orders of ma...