实验基础

Gram-staining is a four part procedure which uses certain dyes to make a bacterial cell stand out against against its background. The specimen should be mounted and fixed on a slide before you procede ...

1. In microfuge tubes spin down 0.1 ml of uninduced cells grown to near saturation or 0.15 ml of IPTG induced cells. Remove YT (or LB) media with a pipetman or drawn out pasteur pipette and freeze cel ...

The 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 sample from being ...

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 ...

Plasmid (pUC series) containing genomic DNA fragments are maintained in E. coli strain DH5aTM. The E. coli cultures are routinely cultured at 37 C on Luria-Bertani (LB) agar on or in LB broth containi ...

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 of culture should b ...

Purpose: Bacterial strains may be stored indefinitely at low temperatures (- 20 degrees C and -80 degrees C) in 15 to 40 glycerol. It is lab policy to prepare a frozen stock of newly acquired or creat ...

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. Add 2 ml (1/100 volu ...

1. Initial inoculum: - 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 c ...

Principle: Bacterial cells are streaked onto a medium to obtain an independent isolate. This is done to reduce the likelihood of working with a culture which has become contaminated and/or has accumul ...

Cloning of PCR products Stocks: LB Agar: Luria Broth after Lennox: per Liter Tryptone 10 g Yeast Extract 5 g Sodium Chloride 5 g Bact. Agar 15 g ...

3 agar (200 ml) Add 6 grams agar to 200 ml deionized water. Autoclave to sterilize. 1.6 agar (200 ml) Add 3.2 grams agar to 200 ml deionized water. Autoclave to sterilize. Alkaline Lysis Solution stoc ...

IntroductionThe term phagocytosis itself describes its mean phage = engulfment; cytosis: cell process. In other words phagocytosis is the cellular process of engulfing solid particles by the cell memb ...

The following protocol has been used successfully to 15N or 13C/15N label our proteins using our pET1120/BL21(DE3) expression system: Preparing M9 minimal media begins with preparing a 5x stock soluti ...

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

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 ...

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 below this temperature as the agar ...

Recipes: 1) LB BrothMake 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 BrothCombine: NaCl ...

AMPICILLIN Beta-lactam-antibiotics are not very stable when dissolved. Slow but steady degradation happens even when frozen to -20°C. Therefore commercial beta-lactam-antibiotics are sold as dried p ...

Grow cells overnight in 500 ml broth medium. Pellet cells by centrifugation and resuspend in 5 ml 50 mM Tris (pH 8.0) 50 mM EDTA. Freeze cell suspension at -20C Add 0.5 ml 250 mM Tris (pH 8.0) 10 mg/m ...