TAILPCRProtocol
TAIL is a series of reactions that are intended to map where a T-DNA (transfer DNA) has inserted within the genome. The main components of the 3 reactions are the AD (Arbitrary Degenerate) primers, border primers, and DNA from the T-DNA lines that are to be mapped. AD primers are degenerate primers that anneal throughout the genome. The border primers are specific for the left and right borders of the T-DNA. From the......阅读全文
TAIL－PCR－Protocol
TAIL is a series of reactions that are intended to map where a T-DNA (transfer DNA) has inserted within the genome. The main components of the 3 react
TAILPCR（thermal－asymmetric－interlaced－PCR)简介
在分子生物学研究中,基因克隆和分子杂交的探针制备等操作常需分离与已知DNA序列邻近的未知序列，TAIL-PCR又叫热不对称交错PCR，能够较好地解决上述难题。该技术通过3个嵌套的特异性引物分别和简并引物组合进行连续的PCR循环，利用不同的退火温度选择性地扩增目标片段，所获得的片段可以直接用做探针标记
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
TAILPCR（Termal－Asymmetric－Interlaced－PCR）基础知识
很早就有用随机引物的PCR，但由于无法有效地控制由随机引物引发的非特异产物的产生，所以一直未能广泛应用。近年来由IJiu等设计的TAIL- PCR(Termal Asymmetric Interlaced PCR)又叫热不对称交错PCR，则解决了这个问题，后来有研究表明，经改良过的TAIL-
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
RTPCR－PROTOCOL
RT-PCR PROTOCOL材料与方法………………………………………………………… 1．材料 ………………………………………………………1.1 供试用组织(细胞)…………………………………1.2 主要仪器设备………………………………………1.3 主要试剂……………………………………………1.
转基因
DNA PreparationGene TransferEmbryo TransferTransgenic IdentificatioinOthersTransgenic Outline (University of Michigan Transgenic Animal Model Core)Thi
Protocol－for－competitive－RTPCR
For quantifying mRNA, we use a competitive RT-PCR protocol with internal standard RNAs. These are added in a defined quantity to the RNA sample prior
SYBR－Green－Quantitative－PCR－Protocol
SummaryQuantitative PCR is a method used to detect relative or absolute gene expression level. All qPCR involves the use of fluorescence to detect the
Single－tube－confirmation－PCR－protocol
The following colony PCR protocol has been designed to be performed in individual reaction tubes. We usually test three colonies from each transformat
DNA抽提
DNA抽提（主要内容如下）· Working with DNA· DNA Extraction from Bacteria and Other Organisms· DNA Extraction from Cell and Tissue· Mitochondria DNA Isola
REALTIME－PCR－WITH－SYBR－GREEN－I－PROTOCOL
1、反应体系 25ulCocktail 20 ul: Primer FP 1 ul + Primer RP 1u + 2 * SYBR GREEN I MIX 12.5 + H2O 5.5 ul cDNA 5 ul2、Primer 浓度，需要摸索，从200nM到700nm等，设置不同浓度。3、每个引
其它PCR方法
· Standard PCR Protocol (Molecular Biology Techniques Manual)The followings are described in detailRecommended Reagent ConcentrationsRecommend
Adiponectin－Replenishment－Ameliorates－ObesityRelated－Hypertension（二）
Methods Animal and Animal Treatment KKAy male mice were purchased from Japan CLEA (Tokyo, Japan). This strain is a cross between black KK fema
Vacuum/Spin－Protocol－for－Tissue－DNA－Extraction
实验概要The E.Z.N.A.® Tissue DNA Kit provides a rapid and easy method for the isolation of genomic DNA for consistent PCR and Southern analysis. Up to 3
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
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
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
RNAi－protocol
siRNA protocolsOur current strategy with siRNA is to synthesis relatively small amounts enzymatically and use these to test for efficiency by western
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
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
NAi－protocol
siRNA protocolsOur current strategy with siRNA is to synthesis relatively small amounts enzymatically and use these to test for efficiency by western
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.
cDNA
· cDNA Synthesis (Crawford Lab)mRNA can be converted into DNA (copy DNA, cDNA) by annealing oligo-dT to the 3' poly-A tail that occurs on
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
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
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
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