RNAiprotocol
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 us......閱讀全文
RNAi－protocol
?siRNA protocolsOur current strategy with siRNA is to synthesis relatively small amounts enzymatically and use these to test for efficiency by western
RNAi
1995年，康乃爾大學的Su Guo博士在試圖阻斷秀麗新小桿線蟲（C. elegans）中的par-1基因時，發現了一個意想不到的現象。她們本是利用反義RNA技術特異性地阻斷上述基因的表達，而同時在對照實驗中給線蟲注射正義RNA（sense RNA）以期觀察到基因表達的增強。但得到的結果是二者都同樣
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
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
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
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
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
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.????????
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總結
RNA干涉（RNAi）是指雙鏈RNA分子使基因表達沉寂的現象，是在線蟲中發現的，在 1998年的一篇Nature論文中被公諸于眾。過去幾年中，科研工作者已明確轉錄后基因沉默現象普遍存在于動、植物中，在機體防御病毒入侵和轉座子沉默效應中起著重要作用。近年來的研究表明，將與mRNA對應的正義RNA和反義
RNAi技術
?DNA芯片檢測siRNA專一性? 長片斷的雙鏈RNA ?(dsRNA)?導入例如植物，真菌，果蠅，線蟲等生物的細胞中會引發同源mRNA的降解——這就是所謂的RNA interference ?(RNAi)。RNAi分兩個步驟，首先是長片斷雙鏈dsRNAs被核酶Dicer切割成21—25個堿基的sm
Transformation－Protocol－for－Arabidopsis
Transformation Protocol for Arabidopsis – Abbreviated Germinate seed in pots ↓ 4 weeks Streak bacteria onto YM/MinA ↓ 2-3 days 28°C Spray/dip ba
Silver－Staining－Protocol
1x 40min - overnight?????50% MeOH, 12% Acetic Acid 1x 30min??????????????????????????????????50% MeOH, 12% Acetic Acid, 0.05% 37% Formaldehyde 3x 2
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 whic
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)
Yale－Immunofluorescence－Protocol
實驗概要We provide a protocol for fixation, immunostaining, and imaging in 384-well Plates.主要試劑Reagents1.?384-well view plates (Aurora)2.?HUVEC (pooled, L
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
BrdU－Labeling－Protocol
實驗概要The thymidine analog, 5-bromo-2-deoxyuridine (BrdU),is a common reagent used for cell proliferation assays and for the detection of apoptotic
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
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
Bacteria－Culture－Protocol
Bacteria Culture ProtocolBy 徐曉政1、TBS Medium Preparation:Prepare 1L of TBS medium contains:Tryptone 12gYeast extract 24gNaCl 5gSodium Succinate 5gGlyce
Protocol－for－Trichl...
實驗概要The ?efficiency of nucleotide incorporation in DNA/RNA polymerization ?reactions (e.g. transcription, reverse transcription, and DNA ?replication)
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
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
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
Histone－blotting－protocol
實驗概要?Western blot detection of histone proteins.?實驗步驟?The ?following protocol refers to the western blot detection of histone ?proteins derived from p
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
Adhesion－Assay－Protocol
Materials to be prepared beforehand:1) Washing Buffer--0.1% BSA in medium (DMEM or RPMI)2) Blocking Buffer--0.5% BSA in medium (DMEM or RPMI)3) Lamini