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erythrophagocytosis assay

OUTLINEThe erythrophagocytosis assay is performed to compare the phagocytic rate with and without anti-eythrocyte antibody in either macrophages from control animals or virus-infected counterparts.PRO ...

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macrophage dichloromethylene diphosphonate)

OUTLINEClodronate (dichloromethylene diphosphonate) is used for treatment of osteolytic diseases and osteoporosis Its effect is due to inhibittion of the function of osteoclasts. This drug is also tak ...

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Enrichment of PBMCs with monocytes

Enrichment of PBMCs with monocytesContributed by Dr.A.GratchevThis protocol is used in our lab to reduce the costs of the cell sorting with MACS reagents. The cell suspension obtained after this proto ...

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peritoneal macrophage preparation

OUTLINEThis modification of peritoneal macrophage preparation is designed for a subsequent thrombophagocytosis test. PROTOCOLSacrifice a mouse (use dry ice CO2) Sink a mouse in the 5% water solution ...

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Phagocytosis

Phagocytosis Fluorescent labeling of yeastResuspend 5 grams of yeast in 50 ml PBS in 100 ml flask Put flask for 30 minutes in boiling water bath while stirring. Wash 5 x with PBS and 2 x in PB. Adjust ...

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Purification of human mononuclear cells and neutrophils

Purification of human mononuclear cells and neutrophilsby Chang-Duk Jun 01/19/2000 PurposeMaterials10ml 6% dextran + 7ml citrate/citric acid Dextran: T500 -- 6g+100ml PBS Citrate solution: 25g Na Citr ...

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Agglutination Assays

Agglutination AssaysREFERENCE: Lanyi B. and T. Bergan. Methods in Microbiology Vol 10: 93-168. BACTERIAL AGGLUTINATION: Bacterial agglutination is performed using either P. aeruginosa cells autoclaved ...

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SMEAR PREPARATION

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 ...

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Bacteria Growth and Culture Bacteria Growth and Culture

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 ...

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Maintenance of Probes in bacteria including Escherichia coli

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 ...

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Basic procedures for bacteria culture

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 ...

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Growing Overnight Cultures

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 ...

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Lyophilizing Cells

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 ...

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Replica Printing

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 ...

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Preparing Overnight Bacterial Culture

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 ...

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Streaking Bacteria for Single Colonies

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 ...

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Agar Plates for Selection of Clones in Bacteria

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 ...

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Expression Protocol in M9 Minimal Media via T7 Promoter

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 ...

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M9 Plate Supplementation Table

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 ...

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Plating in Top Agar

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 ...

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