... 35 μL ddH 2 O Incubate for 4 minutes at 95°C in a PCR machine or in a beaker of boiling water. If using...likely to shear the envelope of your virus. o. Virus may be stored at 4°C for a few days, but should be frozen...miniprep DNA) 5 μL 10x NEB buffer 1 1 μL AgeI add ddH 2 O to bring to 50 μL final volume Incubate at 37°C for...Qiaquick gel extraction kit. Elute in 30 μL of ddH 2 O. Digest eluate with EcoRI. Mix: 30 μL pLKO.1 TRC-cloning...μL 10x NEB buffer for EcoRI 1 μL EcoRI 14 μL ddH 2 O Incubate at 37°C for 2 hours. Run digested DNA on ...Qiaquick gel extraction kit. Elute in 30 μL of ddH 2 O. Measure the DNA concentration. C.4 Ligating and Transforming...DNA ligase buffer 1 μL NEB T4 DNA ligase add ddH 2 O to bring to 20 μL final volume Incubate at 16°C for...

...chloride 12.0 g agar-agar 1 L Sterile H 2 O Sterile plates of your desired size - we usually use 60 mm x 15 mm...indicated, the antibiotic powder can be dissolved in dH 2 O. *Carbenicillin can be used in place of ampicillin...with an absorbent material to reduce condensation. The plates are then stored at 4 ℃ until use. Once your... with resistance to that antibiotic - usually conferred by a plasmid carrying the antibiotic resistance...LB-agar powder. Transfer the LB-agar powder you’ve measured out into an appropriately sized bottle for autoclaving... is necessary to prevent your molten agar from boiling over in the autoclave. Transfer the sterile water...the opening of the bottle with its cap or aluminum foil (but do not make an air-tight seal!) and tape the...

... 4 g NaCl 4 g Tryptone 2 g Yeast Extract and dH 2 O to 400 mL Note: If your lab has pre-mixed LB agar ...the suggested amount, instead of the other dry ingredients above. Media without growth (top) and with growth...cover the entire top of the bottle with aluminum foil. Autoclave and allow to cool to room temperature... Loosely cover the culture with sterile aluminum foil or a cap that is not air tight. Incubate bacterial...