常用设备/其他

Vapor-Phase Sterilization Useful when sterilizing several tubes of seeds. Use microfuge tubes marked with pencil instead of pen (pen will disappear). Poke a small hole into the top of the tube. Put seeds into tubes these tubes. Do not fill tube with seeds past the .1ml line on the tube. Place tubes closed (b ...

I. Prepare plant tissue samples: 1. Weigh ~0.5g fresh leaf tissue (3-5cm2), grind using liquid nitrogen. 2. Add ~0.4ml Plant Extraction Buffer, grind until slurry, transfer to 1.5ml tube.3. Spin down for 15 minutes at 4°C, transfer 0.2-0.3 ml supernatant to fresh tube. 4. Add an equal volume of 2X Laemmli's buff ...

1. Scoop dry soil into an autoclave bin. Fill until the soil is approximately one inch from the top after packing down with the scooper. 2. Cover the bins with aluminum foil and put a piece of autoclave tape on each one. Make sure the outsides of the bins are dirt free before putting in the autoclave. 3. Autoclave on grav ...

(Janine’s protocol)1. Add 200µl of acetone, vortex briefly. 2. Remove acetone and pipette into flat-bottom 96-well plate (polystyrene) 3. Centrifuge plate at 3000 x g for 3-5 minutes. 4. Invert plate carefully and blot dry with paper towels 5. Add 40µl of X-Gluc stain solution to each well (multi-pipett ...

PLANTING SEEDwith tweezers- recommended for pots: 1. Wet small tweezer tips with dH20. 2. Touch one tip to a seed. 3. Move seed to soil by touching tweezer tip to desired spot on soil. 4. Cover pots with humidomes. Let pots sit in 4°C room for 2 days and then move up to growth room. with 0.1% agar- recommended for flats 1. Weigh out s ...

0.01% boric acid 1 mM MgSO4 2mM CaCl2--this was used with media containing GABA. If no other nitrogen source is present, use 1mM CaCl2 and 1 mM CaNO3 18% sucrose pH 6.5 Air dry flowers for 1-2 hours (optional--this helps with release from anthers but is not essential), then dab anthers onto surface of a 25-30µL drop of ge ...

Complementation is performed to rescue the phenotype of a mutant plant with the cloned wildtype gene. This can prove that the gene cloned is in fact the gene that was mutated in the mutant plant. I. Cloning wildtype gene into the plant transformation vector: 1. Design primers for region to be amplified.** ...

Basta SelectionPlant 500 - 1000 plants per flat. (It's a good idea to plant two control pots, one with wild-type and another with a known Basta resistant strain). Apply BASTA solution after the cotyledons have emerged, but before the true leaves have extended. Apply the spray in the fume hood. Wait one day be ...

Fluorescein Diacetate (FDA)1. Place pollen on slide. 2. Incubate in humid chamber (wet Whatman in Tupperware) ~ 30 min. 3. Add drops of FDA (2mg/ml in acetone) to Pollen Growth Media (~/ml) until it becomes turbid. 4. Add 1 drop to pollen. Incubate 10’ 5. Add slide and view under FITC. Pollen Growth Media2X For 4 Liters: ...

Materials Gilson (or Warburg) Respirometer at 37° CMitochondria suspension from Exercise 8.4 Krebs Phosphate Ringers (KPR)10% (w/v) Glucose0.39% (w/v) Sodium azide18.4 mg% (w/v) Dinitrophenol (DNP)6.64% (w/v) Sodium Malonate10% (w/v) KOH Procedure When you enter the lab, check that the ref ...

Aniline Blue Staining of Pollen/Pollen Tubes花粉管染色Materials Fixative: 10% Acetic Acid in ethanol 1M NaOH 50 mM KPO4 buffer, pH 7.5: 4.17 mL 1M K2HPO4 0.83 mL 1M KH2PO4 0.01% aniline blue in 50 mM KPO4 buffer (dye) KPO4 buffer made with 50% glycerol (mounting media) Method 1.Submerge pistil tissue in about 250 ...

Materials 0.01 M Potassium Phosphate Buffer, pH 7.4Sat. K Fe(CN)0.6 M Sodium Malonate (Malonic acid, sodium salt)5 mM KCN0.02% (w/v) p-phenylenediamine oxalate (PPDO) Mitochondria suspension from Spectrophotometer and cuvettes Procedure Prepare a series of 5 tubes as follows: Substan ...

Materials Mitochondrial suspension from Exercise 8.4 Sodium dithionite0.2 M Sorenson Phosphate Buffer, pH 7.51% (w/v) Bovine Serum Albumin 0.005 M Potassium Cyanide0.00025 M Dichlorophenolindophenol0.6 M Sodium succinate, pH 7.50.6 M Sodium malonate, pH 7.50.033% (v/v) Phenazine M ...

Materials Chloroplast suspension from Exercise 8.1AcetoneSpectrophotometer and tubesClinical centrifuge and tubes Procedure Pipet 1.0 ml of a chloroplast suspension into a 15 ml centrifuge tube and add 8.0 ml of acetone. Mix thoroughly. Add 1.0 ml of distilled water, mix again and centr ...

Materials Gilson respirometer equipped with photoflood lampsChloroplast suspension Chloroplast suspension buffer 0.1 M NaHCOInhibitor 1 (1 x 10 DCMU)Inhibitor 2 (5 x 10 DCMU) Procedure In general, use the procedure listed in Exercise 8.7 for mitochondria respiration, except that t ...

Materials Chloroplast suspension 0.035 M NaClBoiling water bathIce bath5 x 10 M 3-(3,4-Dichlorophenyl)-1,1,-Dimethylurea (DCMU) 0.1 M KHPO/KHPO Buffer, pH 6.51 x 10 M Dichlorophenolindophenol (DCPIP) Procedure Dilute the chloroplast suspension with 0.035 M NaCl to yield a final chlor ...

Materials Rat, mouse or suitable source of fresh liver 0.25 M sucrose in 10 mM HEPES buffer, pH 7.5 (Homogenization buffer)0.25 M sucrose in 10 mM HEPES, pH 7.5 + 1 mM EDTA (Suspension buffer)Teflon homogenizerRefrigerated centrifugeJanus Green BHemacytometer and microscope Procedure Sac ...

Materials Fresh spinach leavesGrinding solution0.33 M Sorbitol10 mM Sodium pyrophosphate (NaPO)4 mM MgCl2 mM Ascorbic AcidAdjust pH to 6.5 with HClChopping board and knifeChilled mortar and pestleCheeseclothRefrigerated preparative centrifugeSuspension soluti ...

Transgenic Mouse-Methods and Protocols转基因鼠 实验方法Methods in Molecular Biology VOLUME209Transgenic Mouse-Methods and Protocols转基因鼠 实验方法Edited by Marten H. Hofker, Jan van Deursen2002 Humana Press Inc.PDF格式，362页，9M，点击此处下载。Table of Contents 1. Introduction: The Use of T ...

实验动物解剖学－经典的动物实验方法指导实验动物解剖学南开大学实验动物解剖学编写组高等教育出版社，1988年PDF格式，318页，M，点击此处下载本书内容主要分为上下两篇，共13章。上篇为系统解剖学，选择兔、狗、猫、豚鼠、大白鼠、鸽和蛙等7种常用实验动物分别加以介绍。下篇为局部解剖学，以兔、猫、狗为主要材料，分别介绍了头部、颈部、胸部、腹部、腰背部及股部的局部解剖。此外附录中还介绍了实验动物简介，活体解剖、尸体解剖、血管神经 ...