NAiprotocol
siRNA protocolsOur current strategy with siRNA is to synthesis relatively small amounts enzymatically and use these to test for efficiency by western blotting. This allows us to test several candidate siRNAs quite cheaply. The yields are good enough for reasonable amounts of experimental work, but for larger use we get successful siRNAs synthesised chemically by Dharmacon.DesignThe Tuschl lab siRNA users gu......閱讀全文
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.????????
ELISA－protocol
ELISA protocol：1.取5－10ul BMMY表達上清用0.05M NaHCO3稀釋到100ul鋪ELISA板，37度或室溫振蕩大于1小時。注意一定要做一個GS115空菌株表達上清作為陰性對照，最好還找一個帶有histag的蛋白作為陽性對照。2.TPBS洗板3次，方法：倒掉鋪板液，倒置于
ELISPOT－Protocol
實驗概要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
RNAi－protocol
?siRNA protocolsOur current strategy with siRNA is to synthesis relatively small amounts enzymatically and use these to test for efficiency by western
Immunoprecipitation－Protocol
實驗概要Immunoprecipitation ?is a procedure by which proteins or peptides that react specifically ?with an antibody are removed from solution and examined
ELISPOT－Protocol
實驗概要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
實驗概要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
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
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
Urea－Lysis－Protocol
Urea?lysis?buffer????????????9M Urea, 2.5mM EDTA, 2.5mM EGTA, 1% DTE, 4% CHAPS????????????make 10ml and aliquot 10x1ml, freeze at -70°C?Lysate?prepara
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
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
Cell－Extraction－Protocol
實驗概要Primary tissues ?are valuable tools for the study of intracellular and extracellular ?markers which characterize disease states. We have developed
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
Transformation－Protocol－for－Arabidopsis
Transformation Protocol for Arabidopsis – AbbreviatedGerminate seed in pots↓ 4 weeksStreak bacteria onto YM/MinA↓ 2-3 days 28°CSpray/dip bacterial sus
Cytotoxicity－Assays－Protocol
Cytotoxicity Assays ProtocolCell-mediated cytotoxicity was determined by using a standard microcytotoxicity assay. Briefly, target cells were pelleted
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
Protocol－for－dsRNA－Synthesis
實驗概要? ? ? ? We routinely produce dsRNA by in vitro transcription of a PCR generated DNA template containing the T7 promoter sequence on both ends
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
Sandwich－ELISA－Protocol
實驗概要The ?Sandwich ELISA measures the amount of antigen between two layers of ?antibodies (i.e. capture and detection antibody). The antigen to be ?mea
Protocol－of－Northern－blot
Protocol of Northern blot質粒的轉化和擴增質粒的鑒定目的基因片段的切割3.1樣品雙酶切(175μl水解體系)DW 115μlBuffer B(10×) 17.5μlBaM H 15μlPst I 17μlDNA(MMP-9) 16μlBSA 4.5μl37℃水浴，3h。3.2
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
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
Western－Blot－Protocol
一、提取抗原蛋白將提取RNA途中留存的樣品，加入150μl 100%酒精充分混勻，靜置5min(RT), 2000×g , 4℃離心5min,?吸取上清至新管中,?加入750μl異丙醇,?混勻,?靜置10min(RT), 12000×g, 4℃離心10min,?棄上清,?加入1ml 0.3mol/L
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
Immunofluorescence－Microscopy－Protocol
實驗概要Immunofluorescence ?allows the imaging of a specific factor in cells or tissue sections ?through the use of a specific antibody chemically which i
Dot－blot－protocol
實驗概要A ?technique for detecting, analyzing, and identifying proteins, similar ?to the western blot technique but differing in that protein samples are