LentiVirusProtocol
Lenti-Virus Protocol张端午A. For packing virus in 6-well plate: (if in 12-well plate, reduce all to 1/2)1. Mix the following plasmids in a 1.5 ml eppendorf tube,1.5 μg Lenti-vector + 1.5 μg packing plasmids = 3 μg total0.5:0.3:0.2 (pMDL =0.75 μg, VSV-G=0.45 μg, REV=0.3 μg)add 200 μl OPTI-MEM, mix well.2. Mix 7.5 μl LF2K in 200 μl OPTI-MEM, incubate for 5 min.3. Mix the LF2K mixture with plasmids mixture, incubate for 15......阅读全文
LentiVirus－Protocol
Lenti-Virus Protocol张端午A. For packing virus in 6-well plate: (if in 12-well plate, reduce all to 1/2)1. Mix the following plasmids in a 1.5 ml eppendo
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
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
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
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
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
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
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.
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
Cell－Extraction－Protocol
实验概要Primary tissues are valuable tools for the study of intracellular and extracellular markers which characterize disease states. We have developed
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
Phycoerythrin－conjugation－protocol
Phycoerythrin conjugation protocolDavid's method modified from references (2) and (3). I used this method to conjugate a mouse IgG2a monoclonal an
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
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
Dot－Blot－Protocol
a. Label nitrocellulose membrane (using a pencil) to identify protein elution fractions.b . Pipette 2μl from each fraction onto the membrane, allow th
Protocol－for－dsRNA－Synthesis
实验概要 We routinely produce dsRNA by in vitro transcription of a PCR generated DNA template containing the T7 promoter sequence on both ends
Migration－Assay－Protocol
Materials to be prepared beforehand:1) FBS free medium2) 10% FBS medium3) Cell migration filter insert ( Transwell®, 12mm Diameter, 12 μm Pore Size.)P
RNA－Isolation－Protocol
Stabilize RNAStart with 15 ml E. coli Culture containing 7.5* 109 cells (OD600= 0.2 Dilute cells or scale up)Pipet 30 ml of RNAProtect Bacteria Reagen
Protocol－for－Cell－Fusion
Healthy Sp2/0 cells should be rapidly growing by this time. Sp 2/0 cells should be started about two weeks before the cell fusion. Every two days, the
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
Western－Blot－Protocol
一、提取抗原蛋白将提取RNA途中留存的样品，加入150μl 100%酒精充分混匀，静置5min(RT), 2000×g , 4℃离心5min, 吸取上清至新管中, 加入750μl异丙醇, 混匀, 静置10min(RT), 12000×g, 4℃离心10min, 弃上清, 加入1ml 0.3mol/L
Nucleolar－Isolation－Protocol
We recommend that you first download and read this page as a PDF file. Using that as your guide, you can then follow the protocol below and view a Qui
Protocol－of－Northern－blot
Protocol of Northern blot质粒的转化和扩增质粒的鉴定目的基因片段的切割3.1样品双酶切(175μl水解体系)DW 115μlBuffer B(10×) 17.5μlBaM H 15μlPst I 17μlDNA(MMP-9) 16μlBSA 4.5μl37℃水浴，3h。3.2
Protocol－for－Trichl...
实验概要The efficiency of nucleotide incorporation in DNA/RNA polymerization reactions (e.g. transcription, reverse transcription, and DNA replication)
Dot－blot－protocol
实验概要A technique for detecting, analyzing, and identifying proteins, similar to the western blot technique but differing in that protein samples are
Sandwich－ELISA－Protocol
实验概要The Sandwich ELISA measures the amount of antigen between two layers of antibodies (i.e. capture and detection antibody). The antigen to be mea
cDNA/AFLP－Protocol
Preparation of Para-magnetic beads from Promega cat#Z5482:a) suspend magnetic particles in bottle - transfer 200 ul (200 ug) of beads per sampleof RNA
RNA－Isolation－Protocol
RNA Isolation Protocol(Revised 5-15-2003)Stabilize RNAStart with 15 ml E. coli Culture containing 7.5* 109 cells (OD600= 0.2 Dilute cells or scale up)