抗体抗原实验

SMEAR PREPARATIONThe preparation of a smear is required for many laboratory procedures including the Gram-stain. The purpose of making a smear is to fix the bacteria onto the slide and to prevent the ...

Flagella and motility monotrichous flagella - the bacterial cell has a single flagella peritrichous flagella - the bacterial cell has several flagella which are located at various sites on the cell su ...

Maintenance of Probes in bacteria including Escherichia coliPlasmid (pUC series) containing genomic DNA fragments are maintained in E. coli strain DH5aTM. The E. coli cultures are routinely cultured a ...

A. Phenol extraction of DNA samplesPhenol extraction is a common technique used to purify a DNA sample (1). Typically an equal volume of TE-saturated phenol is added to an aqueous DNA sample in a micr ...

Jeff Lawrence (M02)1. Place 2 ml of the appropriate sterile medium in a 13 mm yellow-capped culture tube. If more culture is needed place up to 5 ml in a 16 mm green-capped culture tube. Large amount ...

Lyophilizing Cells Inoculate 200 ml L-broth supplemented with appropriate antibiotics with the bacteria to be lyophilized. Incubate the culture at 37°C with vigorous shaking (200-250 rpm) overnight. A ...

Jeff Lawrence (M05)1. Sterilize Velvet Squares with widths at least 50% longer than the diameter of a petri plate.2. Streak strains to be printed for single colonies.3. Prepare a printing Master Plate ...

Materials: Sterile LB medium (Luria-Bertani Medium) with or without antibiotic: water 500 ml bacto-tryptone 10 g bacto-yeast extracts 5 g sodium chloride 10 g stir to dissolve pH to 7.0 autoclave to s ...

Jeff Lawrence (M01)1. - From solid media touch an isolated colonies with a sterile applicator or toothpick.- From liquid media touch the culture with the end of a sterile applicator.- From a frozen cu ...

Agar Plates for Selection of Clones in BacteriaCloning of PCR productsStocks:LB Agar: Luria Broth after Lennox:per LiterTryptone10 gYeast Extract5 gSodium Chloride5 gBact. Agar15 gpH 7.0Autoclave ...

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 ...