Media and buffers recipes Miniprep buffers P1 (resuspension buffer) P2 (lysis buffer) N3 (neutralization buffer) P3 (neutralization buffer) PB (PCR cleanup or EnoA+ removal) PE (wash buffer) QG (gel extraction) Terific Broth (TB) YPAD Drop-out media 10X HBS Back to Protocols Terific Broth (TB) For 1 L of media: 47.6 TB mix 4 ml glycerol Add water to 1 L and autoclave at 121°C for 15–20 minutes. After autoclave add: 0.1% - 1% glucose 10 mM MgSO4 antibiotics YPAD For 1 L of media: 10 gr Yeast extract 20 gr Peptone 80 mg Adenine sulfate 20 gr Dextrose (Glucose) For plates add: 20 gr Agar yeast drop-out media For 1 L of media: 1.92 gr drop-out mix 6.7 gr yeast nitrogen base w/o AA 20 gr carbohydrate (glucose/galactose) For plates add: 20 gr Agar P1 (resuspension buffer) Keep refrigiratedShake well before using 50 mM Tris-HCl pH 8.0 10 mM EDTA 100 µg/ml RNase A 43 µg/ml Thymophthalein (blue pH indicator) 1000X Blue pH indicator dye: 43 mg/ml Thymophthalein in 100% ethanol P2 (lysis buffer) 200 mM NaOH 1% SDS N3 (neutralization buffer) Neutralization buffer for spin columns 4.2M guanidine hydrochloride (GuHCl) 0.9M potassium acetate pH 4.8 P3 (neutralization buffer) Neutralization buffer for bacmid purification, midi, maxi, giga prepsDo not use with spin columns! 3.0 M potassium acetate pH 5.5 PB buffer extra wash step for EndA+ removal or PCR purification 5M guanidine hydrochloride (Gu-HCL) 30% isopropanol QG (gel extraction buffer) 5.5 M guanidine thiocyanate (GuSCN) 20 mM Tris-HCl, pH 6.6 (25ºC) PE (wash buffer) 80% ethanol 10mM Tris-HCl pH 7.5 (25ºC) 10X HEPES buffered saline (HBS) 200 mM HEPES pH 7.5 1.5 M NaCl