LentiVirusProtocol
Lenti-Virus Protocol張端午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......閱讀全文
LentiVirus－Protocol
Lenti-Virus Protocol張端午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
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
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
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
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.????????
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
NAi－protocol
siRNA protocolsOur current strategy with siRNA is to synthesis relatively small amounts enzymatically and use these to test for efficiency by western
RNAi－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
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
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
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
Silver－Staining－Protocol
1x 40min - overnight?????50% MeOH, 12% Acetic Acid1x 30min??????????????????????????????????50% MeOH, 12% Acetic Acid, 0.05% 37% Formaldehyde3x 20min?
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
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
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)
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
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
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
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
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)
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
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
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
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