lb agar plates kanamycin protocol for humans

How to Make Agar Plates

Agar is the gelatinous substance that sits inside the petri dishes used by scientists and students alike Agar is the perfect substance for biological experiments as it holds up to bacteria and doesn't disintegrate easily There are a number of ways to make an agar plate or agar-filled petri dish You can purchase

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Preparation of LB liquid medium

Preparation of LB agar plates: (~30-35 plates) Prepare 1L of LB liquid (25g broth powder per 1L) in a 2000mL Erlenmeyer flask Add 15g (1 5%) bacteriological agar Swirl to mix Powder will not dissolve completely that is ok Add a fresh piece of aluminum foil to cover the top and add a fresh piece of autoclave tape

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Pouring LB

Pouring LB-Agar+Antibiotic Plates This recipe is for 500 mL of LB agar This makes about 20 10-cm plates (1 bag) 5 g bacto tryptone 2 5 g yeast extract 5 g NaCl 7 5 g bacto agar 500 mL Milli-Q water -OR- Materials you need: • LB Agar Miller (Fisher catalog #: BP1425-500) o 40 grams of LB Agar per 1L of MilliQ water • 2000mL Erlenmeyer flask

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How To Make LB Agar Plates

How to make LB agar plates Weigh out 7 5 g agar 5 g tryptone 5 g sodium chloride (NaCl) and 2 5 g yeast extract and add to a 1 L Duran bottle Measure out approximately 400 mL of distilled water and add to the Duran bottle Shake the bottle to dissolve the reagents

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Kanamycin

Kanamycin agar plates isopropyl β-d-1-thiogalactopyranoside (IPTG) lysozyme Terrific broth modified EZmix powder (Sigma) glycerol Tris base Tris maleate sodium chloride imidazole calcium chloride dithiothreitol (DTT) phenylmethylsulfonyl fluoride (PMSF) 2 Laemmli sample buffer (Bio-Rad Catalog #1610737) electrophoresis buffer and Avicel (PH101 Fluka Buchs

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MAKING AND POURING LB AGAR PLATES (STERILE)

MAKING AND POURING LB AGAR PLATES (STERILE) (1 batch makes about 40 plates adjust as needed) Est Total Time: 1-2 Hours Summary: Bacteria need nutrient plates to grow on This protocol will show you how to make selective (with antibiotics) and nonselective (without antibiotics) plates SAFETY

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Media Preparation: Making agar plates containing

NB: LB and L-agar MUST be autoclaved on the same day they are made up Make sure you put autoclave tape on the bottles before autoclaving L-Agar 5g Tryptone 2 5 g Yeast extract 2 5 g NaCl 1 5 % Agar (7 5g in 500 ml) Distilled water to 500 ml The agar will not dissolve until autoclaved Autoclave for 30-45 minutes 1

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Barrick Lab :: ProtocolsAntibioticStockSolutions

Ampicillin degrades quickly in both plates and stock solutions recommended Kanamycin plates only last a few weeks at 4C best to make fresh Kan plates every 2 - 4 weeks Nalidixic Acid : Use at ten-fold higher concentration in motility agar (0 5%) Working concentrations are for liquid or solid media

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Preparing Antibiotics Stock Solution and Ampicillin

I also observed impaired activity of Kanamycin after several months at -20C! 50 mg/ml is suitable concentration for stocks Add 200 l to 100 ml LB-Agar/LB-Medium (100 g/ml) 200 g/ml for Ampicillin works also fine Preparation of Agar Plates Agar Recipes: 10 g of NaCl 10 g of Tryptone 5 g of Yeast Extract 20 g of Agar

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Team:Purdue/Protocol

How to make LB plates plus antibiotics: Follow the recipe card in box for making LB plates being sure to add the agar After autoclaving and when the agar has cooled enough that it's not too hot to touch (about 1 to 1 5hrs) add antibiotics as follows: Ampicillin – add 1ml ampicillin (at 100mg/ml) per liter of agar to obtain a final

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Agar Plates and Media Protocol

Kanamycin 50 g/L - Green Ampicillin 100 g/L - Red For every mL of LB agar solution add 1 L of antibiotics Since I was using 300 mL of water I added 3 mL of kanamycin The plates are poured and cooled then marked with a marker to signify the antibiotic Agar stabs are from the same protocol just poured into tubes and capped

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LB (plus Antibiotic) Agar Plates Preparation

LB + Kanamycin - (optional black band) single vertical green band 11 When it's cooled down enough add the appropriate amount of antibiotic(s) to the medium and swirl to mix: 100ug/mL Ampicillin 34ug/mL Chloramphenicol 10ug/mL Kanamycin 12 You can pour the LB agar from the flask into a sterile 500-mL beaker for easier transfer onto the plates

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A Make Luria broth (LB) and LB agar plates —

When the agar has spread to cover about two thirds of the dish stop pouring and the agar should spread to cover the entire plate You may need to tilt the plate slightly to get the agar to spread out completely If you pour in too much the plate will be fine but it will reduce the number of plates you can make per batch

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Making Agar Plates for Bacterial Growth

Petri Plates LB Agar Media Scale Glassware To start we will talk about a bacterial base in which we use LB AGAR A common ratio to remember when making your LB AGAR mix is 40g to 1L of water ratio This ratio will make about 80 plates This protocol is going to walk you through making 10 plates

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Preparative steps: culture media agar plates antibiotics

4 Preparative steps: culture media agar plates antibiotics and glycerol 3 Preparation Antibiotics 3 1 Kanamycin 30 mg/mL stock solution in H 2 O Final concentration in cultures and LB-agar: 30 g/mL Filter stock solution using 0 2 m filter 3 2 Chloramphenicol (for protein expression) 25 mg/mL stock solution in ethanol

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How to Make Agar Plates

Agar is the gelatinous substance that sits inside the petri dishes used by scientists and students alike Agar is the perfect substance for biological experiments as it holds up to bacteria and doesn't disintegrate easily There are a number of ways to make an agar plate or agar-filled petri dish You can purchase

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BD BBL™ CDC Anaerobe Laked Sheep Blood Agar with Kanamycin

L007360 2 of 4 BD BBL CDC Anaerobe Laked Sheep Blood Agar with Kanamycin and Vancomycin was formulated by Dowell et al of the Centers for Disease Control and Prevention as an enriched selective medium for the isolation and cultivation of Prevotella melaninogenica Fusobacterium necrophorum Fusobacterium nucleatum and other fastidious obligately

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