GeneralCloningProtocols
Large Scale Preps: (See Large scale plsasmid prep protocol for more details)Cultures: Inoculate a 5 mL LB/Amp (50 - 100 µg/mL) culture in early a.m. with a single colony. Use all 5 mL to inoculate a 500 mL LB/Amp culture in the evening. Alternatively the 5 mL culture can also be set up as an overnight culture.Save 1 mL for a glycerol stock if necessary (see step 6c, below). Prepare remainder according to ......阅读全文
Rat－Blood－Collection－Protocols
实验概要The procedure presented below describes a method for collecting rat blood.实验步骤Rat should be fully anesthetized (e.g., unresponsive to toe pinch).1
Protocols－for－LCM－preparation－and－analysis
Protocols for LCM preparation and analysis I. Preparation, LCM and RNA/DNA extraction of Frozen Tissue SectionsA. EmbeddingB. CuttingC. StainingII. Pr
ORNL－MICROARRAY－HYBRIDIZATION－PROTOCOLS
Direct labeling of total RNA with Cy3 and Cy5:A. MATERIALSRNeasy® Mini Kit (Qiagen; Cat # 74106) SuperScript II RT (200U/µL) (Life Technologies; Cat #
Streptomyces:Protocols/Spore－Prep
Spore Prep - Inoculating & HarvestingDescription A spore prep is a method of preserving a sporulating strain of Streptomyces. The stock is stored in 2
General－Laboratory－Procedures,－Equipment－Use,－and－Safety－Considerations
A. Storage .The following properties of reagents and conditions are important considerations in processing and storing DNA and RNA. Heavy metals promo
Cloning－PCR－products－using－TA－vectors
Cloning PCR products using TA vectorsby Paul N. Hengen, Ph.D. *Methods and reagents is a unique monthly column that highlights current discussions in
Western－杂交
Western 杂交（主要内容如下）Preparing of Protein LysatesWestern BlottingFar Western BlottingSemi Dry BlottingStripping MembranesTrouble Shooting and OthersPrepa
DataONE:Protocols/Find－GEO－reuses
Identify reuses of GEO datasetsAimThe aim of this protocol is to collect data on the reuses of datasets in the published literature. This particular p
Red－Blood－Cell－Lysis－Protocols
实验概要BioLegend’s Red Blood Cell (RBC) Lysis Buffer (Cat. No. 420301) has been designed, formulated, and tested to ensure optimal lysis of RBCs in sin
FISH－protocols－for－Drosophila2
3. Methods3.1 RNA Probe Preparation1. Different strategies can be used to prepare template DNA for synthesizing antisense RNA probes by in vitro tr
FISH－protocols－for－Drosophila1
.1 RNA Probe Preparation (see Note 1)1. 1.5 mL microcentrifuge tubes or standard 96-well V-bottom microplates.2. RNAse free water.3. T7, T3 or S
集锦－|－NG－General系列氮气发生器
今年4月，普敦科技将氮气发生器产品线进行了全面的升级，新款NG General系列已正式上线。 目前，普敦氮气发生器拥有【膜分离技术】和【分子筛技术】的一体式/分体式氮气发生器，可以供各品牌的LC-MS、氮吹仪、ELSD/CAD、液氮发生器等设备用气。 一体式 氮气发生器 01【膜分离】
Standard－Protocols－Autoradiography－(35S)
Remove Kodak NTB2 nuclear emulsion from fridge and place at 42oC for around 30-60 mins (until melted).Make up the developer and the fixer and place in
Streptomyces:Protocols/MiniMaxi－Prep
Small Scale Plasmid Isolation (Mini / Maxi Prep)Description A mini prep / maxi prep is used to isolate plasmid or cosmid DNA from bacteria, normally E
PCR基本实验方法（五）
Cloning PCR ProductsT-A Cloning Strategy: Taq and other polymerases seem to have a terminal transferase activity which results in the non-templated ad
PCR基本实验方法（五）
Cloning PCR ProductsT-A Cloning Strategy: Taq and other polymerases seem to have a terminal transferase activity which results in the non-templated ad
基因克隆技术（Gene－Cloning－Techniques）2
二、目的基因和载体的连接获得目的基因后必须将其放在一定的载体内才能在宿主细胞内扩增或表达。目的基因与载体的连接及其后续的转化过程习惯上称为克隆（cloning）。由于目前很多基因都是利用PCR技术获得，因此这里先介绍PCR产物的克隆策略，然后再介绍其他的克隆方式。（一）PCR产物的克隆策略获得PCR
Cosmid－Cloning:－Cell－preparation,－DNA－packaging,－and－Cell－Transfection
Cosmid Cloning: Cell preparation, DNA packaging, and Cell TransfectionProtocol taken from Stratagene's Gigapack packaging extracts instruction man
基因克隆技术（Gene－Cloning－Techniques）2
二、目的基因和载体的连接 获得目的基因后必须将其放在一定的载体内才能在宿主细胞内扩增或表达。目的基因与载体的连接及其后续的转化过程习惯上称为克隆（cloning）。由于目前很多基因都是利用PCR技术获得，因此这里先介绍PCR产物的克隆策略，然后再介绍其他的克隆方式。 （一
信号传导
Cytokine Bioassays (eBioscience)Biological activity of cytokines and their concentrations are commonly measured by cellular proliferation of primary c
信号传导
Cytokine Bioassays (eBioscience)Biological activity of cytokines and their concentrations are commonly measured by cellular proliferation of primary c
Slice－and－Explant－Culture－Protocols－–－Hevner－lab－2002
for axon tracing & cell culture studies in vitro using embyros age E11.5 – E16.5modified from Rubenstein lab and Price lab protocols1. Setupa. 2 hr be
GenomeWide－Identification－of－Transcription－FactorBinding－Sites－in...
Genome-Wide Identification of Transcription Factor-Binding Sites in Plants Using Chromatin Immunoprecipitation Followed by Microarray (ChIP-chip)
CDNA文库
CDNA文库（主要内容如下）· Construction of cDNA Library· Construction of Genome DNA Library· Library Screening OthersConstruction of cD
Infusion－biobrick－assembly
OverviewThis is a method to assemble two BioBricks using the Clontech In-Fusion PCR Cloning Kit and maintains BioBrick standard formats. There are cur
Human－Embryonic－Stem－(ES)－Cell－Protocols——Embryonic－Bodies
Let human ES cells grow until the colonies are large and the cells are pretty piled up - about the time when you would normally split or even a day pa
Human－Embryonic－Stem－(ES)－Cell－Protocols——Media－and－Reagents
Serum Free Media for human ES cells on MEFs: can last for 7-10 daysFinal ConcentrationAmount for 250ml Stock solution80% DMEM-F12200ml20% KO Serum Rep
重组DNA的分离、克隆与测序实验手册
edited by Bruce A. Roe Judy S. Crabtreeand Akbar S. KhanDepartment of Chemistry and Biochemistry The Uni
Cloning－of－small－RNAs－with－5’－phosphate－and－3’－OH－ends3
Load your precipitated PCR samples into 2 consecutive lanes so as not to overload the lanes. For each different sample, I would run a separate ladder
Cloning－of－small－RNAs－with－5’－phosphate－and－3’－OH－ends2
3’ Adaptor Ligation and PurificationHeat shock the RNA by putting at 90°C for 30 seconds. Snap cool on ice.Set up the 3’Adaptor ligation reaction in a