免疫学技术

Expression Protocol in M9 Minimal Media via T7 PromoterThe following protocol has been used successfully to 15N or 13C/15N label our proteins using our pET1120/BL21(DE3) expression system: Preparing M ...

M9 Plate Supplementation TableAmino AcidStockAmount To Spread(per 25 ml plate)URA1 mg/ml280 µlLEU20 mg/ml98 µlMET2 mg/ml500 µlHIS10 mg/ml39 µlTRP10 mg/ml51 µl ...

1. Warm plates to room temperature before use. Cold plates causes the top agar to solidify irregularly. DO not warm plates to 37 C as the top agar will take forever to solidify.2. Prepare Top Agar as ...

Preparation of Agar plates Prepare media and add 1.5 % agar before autoclaving it (15g per liter). After autoclavation cool the media in a 55 degree waterbath. Do not allow the solution to cool bel ...

1. For rich media weigh out appropriate ingredients and place into a flask. Add water until appropriate volume. Use a flask at least 2 times larger than the media volume.2. For minimal medium make sep ...

Preparation of Broth and Plates etc.Recipes: 1) LB Broth Make 16 gm of LB Broth Base (Gibco #M27800C) up to 800 ml in ddH2O. Swirl to dissolve then add 110 µl of 10 N NaOH. Autoclave.2) NZY Broth Comb ...

Transposon-mediated Mutagenesis Step 1 - Amplify ORF from MG16551.0 ul genomic DNA (30ng/ul)5.0 ul gene specific primer mix4.0 ul 2.5 mM dNTP5.0 ul 10x ...

Chromosomal DNA IsolationMETHOD:Grow cells in 2-5 ml broth to late log phase.Pellet 1-2 ml cells in microfuge.Resuspend cells in 400 µl TES (50 mM Tris-HCl 10 mM NaCl 10 mM EDTA pH7.5).Add: 17 µl 30% ...

Triton-Prep Method for bacterial DNA PurificationGrow 5 ( large scale-15ml culture). Harvest in single eppendorf tube (or 15 ml disposable tube). Resuspend pellet with 300ul STET buffer (900ul). After ...

Chromosomal DNA Extraction from Gram-positive BacteriaThis procedure was originally developed for Listeria monocytogenes but has worked well with other Gram+ bacteria we've tried.Pellet cells from 10 ...

RNA Isolation Protocol(Revised 5-15-2003) Stabilize RNAStart with 15 ml E. coli Culture containing 7.undefined 109 cells (OD600= 0.2 Dilute cells or scale up)Pipet 30 ml of RNAProtect Bacteria Reagent (Qiage ...

Isolation of genomic DNA from bacteriaNote: This procedure does not work well with Gram + cocci. Transfer 1.5 mL overnight culture to a 1.5 mL microfuge tube centrifuge for 30 sec decant supernatant.R ...

Preparation of Genomic DNA from Bacteria- using Phase Lock GelTM (Modified from Experimental Techniques in Bacterial Genetics Jones and Bartlet 1990)Materials: TE buffer10% (w/v) sodium dodecyl ...

Isolation and Quantification of Genomic DNA from Mycobacterium tuberculosisThe following protocol describes a "DNA mini-prep" procedure (Part A) for the isolation of chromosomal DNA from Mycobacterium ...

ISOLATION OF RNA FROM BACTEROIDS3 g nodules (fresh or frozen in liquid N2) were ground to a powder in mortar and pestle with liquid N2. To the powder was added ice cold 0.5 M mannitol (or sucrose) 0.0 ...

Purifying Large E. coli Restriction Fragments from Pulsed-Field Gels DNA Preparation E. coli chromosomal DNA is prepared following the method of Heath et al. ( J. Bacteriol. 174 1992). Cells are embed ...

Creation and use of your infectious vector:Plate 5 x 105 293T cells in 6 cm2 dishes containing 5 ml of media. (This can be scaled up if desired). The following day set up (use polypropylene tubes for ...

Some basic Neurospora methods compiled at Stanford Vogel's Minimal Medium D. D. Perkins - Hints for the care feeding and breeding of Neurospora Strain preservation at the FGSC Toxic concentrations of ...

Hints and precautions for the care feeding and breeding of NeurosporaSome of these notes describe what may be established routines in older laboratories but are either unpublished or published in inac ...

Assessment of Magnaporthe grisea mating type by spore PCRJin-Rong Xu and John E. Hamer - Department of Biological Sciences Purdue University West Lafayette IN 47907Isolation of DNA from filamentous fu ...