TransformationProtocolforArabidopsis
Transformation Protocol for Arabidopsis – AbbreviatedGerminate seed in pots↓ 4 weeksStreak bacteria onto YM/MinA↓ 2-3 days 28°CSpray/dip bacterial suspension onto plants↓ 1 day in boxLet plants set seed↓ 3-4 weeksPlate seed to GM + selection↓Transfer transformed plants to the glasshouse Plant material: • Thinly sow Arabidopsis thaliana cv C24 seed into soil. Grow at 20°C under......阅读全文
Immunoprecipitation－Protocol
实验概要Immunoprecipitation is a procedure by which proteins or peptides that react specifically with an antibody are removed from solution and examined
ELISPOT－Protocol
实验概要The Elispot (Enzyme Linked Immuno-Spot) assay provides an effective method of measuring the antibody or cytokine production of immune cells on t
ELISPOT－Protocol
实验概要The Elispot (Enzyme Linked Immuno-Spot) assay provides an effective method of measuring the antibody or cytokine production of immune cells on t
ELISA－protocol
ELISA protocol：1.取5－10ul BMMY表达上清用0.05M NaHCO3稀释到100ul铺ELISA板，37度或室温振荡大于1小时。注意一定要做一个GS115空菌株表达上清作为阴性对照，最好还找一个带有histag的蛋白作为阳性对照。2.TPBS洗板3次，方法：倒掉铺板液，倒置于
Immunoblot－Protocol
This method was successful in our lab using prostate tissue and for our specific objectives. Investigators must be aware that they will need to tailor
RNAi－protocol
siRNA protocolsOur current strategy with siRNA is to synthesis relatively small amounts enzymatically and use these to test for efficiency by western
NAi－protocol
siRNA protocolsOur current strategy with siRNA is to synthesis relatively small amounts enzymatically and use these to test for efficiency by western
ELISPOT－protocol
实验概要The procedure below is a general guideline procedure for ELISPOT. Abcam ELISPOT kits have been designed for detection of various cytokines and g
PCR－protocol
PCR reactionProtocol for 50µl reaction - adjust amounts if necessary, for a 20µl reaction use the same volumes of primer and dNTP-mix, but adjust the
RLGS－protocol
A. Preparation of DNA SolutionIn the case of rice, for example This method may be appllicable for many grass species and some other plants.
Nicotiana－(Nicotiana－tobaccum,－Nicotiana－benthamiana)
Agrobacterium -mediated transformation of Nicotiana species, namely, Nicotiana tobaccum and Nicotiana benthamiana , using leaf disks as the target e
PEGMediated－Protoplast－Transformation－with－Naked－DNA
Direct introduction of DNA into plant protoplasts facilitates a rapid analysis of transient gene expression, as well as the generation of stably t
细菌转化（bacterial－transformation）原理和操作
1．目的学会质粒DNA转化感受态受体菌的技术。2．原理质粒DNA粘附在细菌细胞表面，经过42°C短时间的热击处理，促进吸收DNA。然后在非选择培养基中培养一代，待质粒上所带的抗菌素基因表达，就可以在含抗菌素的培养基中生长。3．器材旋涡混合器，微量移液取样器，移液器吸头，1.5ml 微量离心管，双面微
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
Bacterial－Colony－PCR
Bacterial Colony PCRObjective:This protocol allows rapid detection of transformation success when primers are available to allow determination of corr
Global－Expression－Profiling－of－RNA－from－Laser－Microdissected－Cells－at－...
Global expression profiling of RNA isolated from laser microdissected cells allows one to profile a specific set of cells allowing for enhanced se
DNA克隆
DNA克隆（主要内容如下）· General Procedure· PCR Cloning· Subcloning· ET Cloning· Vector Preparation· Ligation Re
Microarray－Hybridization－Protocol
Introduction:One microarray set consists of 7 nylon membranes with 2.5 x 7.5 cm dimension. 2304 genes were spotted onto nylon membranes (Schleicher an
Nuclear－Extraction－Protocol
实验概要The procedure presented below describes a method for extracting nuclear from several cell lines of human origin.主要试剂Hypotonic Buffer Solution20 mM
Immunofluorescence－Microscopy－Protocol
实验概要Immunofluorescence allows the imaging of a specific factor in cells or tissue sections through the use of a specific antibody chemically which i
Yale－Immunofluorescence－Protocol
实验概要We provide a protocol for fixation, immunostaining, and imaging in 384-well Plates.主要试剂Reagents1. 384-well view plates (Aurora)2. HUVEC (pooled, L
Bacteria－Culture－Protocol
Bacteria Culture ProtocolBy 徐晓政1、TBS Medium Preparation:Prepare 1L of TBS medium contains:Tryptone 12gYeast extract 24gNaCl 5gSodium Succinate 5gGlyce
Intracellular－Staining－Protocol
1. Fix cells- Add16% formaldehyde directly into culture medium to obtain a final concentration of 1.5% formaldehyde.2. Incubate in fixative for 10 min
Histone－blotting－protocol
实验概要 Western blot detection of histone proteins. 实验步骤 The following protocol refers to the western blot detection of histone proteins derived from p
BrdU－Labeling－Protocol
实验概要The thymidine analog, 5-bromo-2-deoxyuridine (BrdU),is a common reagent used for cell proliferation assays and for the detection of apoptotic
Cytotoxicity－Assays－Protocol
Cytotoxicity Assays ProtocolCell-mediated cytotoxicity was determined by using a standard microcytotoxicity assay. Briefly, target cells were pelleted
Western－Blotting－Protocol
实验概要The western blot (sometimes called the protein immunoblot) is a widely used analytical technique used to detect specific proteins in the given s
Tissue－Harvest－Protocol
TISSUES TO BE PROCURED(minimally and preferably within 5-8 hours after death):1. Brain2. Liver3. Muscle4. Skin5: Others as the specific case dictatesP
Colony－PCR－Protocol
1. Pull out eight glycerol stock plates from the –80oC freezer and set on bench top to thaw. Be sure to remove the foil seal before leaving the plates
Silver－Staining－Protocol
1x 40min - overnight 50% MeOH, 12% Acetic Acid1x 30min 50% MeOH, 12% Acetic Acid, 0.05% 37% Formaldehyde3x 20min