Buffer recipes

TAE (1X)

running buffer · per 1000 mL

Agarose gel electrophoresis running/casting buffer.

• Tris base: 4.84 g
• Glacial acetic acid: 1.14 mL
• 0.5 M EDTA, pH 8.0: 2 mL

TBE (1X)

running buffer · per 1000 mL

Higher-buffering-capacity alternative to TAE, common for smaller fragments/PAGE.

• Tris base: 10.8 g
• Boric acid: 5.5 g
• 0.5 M EDTA, pH 8.0: 4 mL

PBS (1X), pH 7.4

wash buffer · per 1000 mL

Phosphate-buffered saline — washes, dilutions, cell culture rinses.

• NaCl: 8 g
• KCl: 0.2 g
• Na₂HPO₄: 1.44 g
• KH₂PO₄: 0.24 g

RIPA lysis buffer

lysis buffer · per 100 mL

Whole-cell/tissue lysis for total protein extraction.

• Tris base (50 mM, pH 8.0): 0.61 g
• NaCl (150 mM): 0.88 g
• NP-40 / IGEPAL CA-630 (1%): 1 mL
• Sodium deoxycholate (0.5%): 0.5 g
• SDS (0.1%): 0.1 g

LB Broth (Miller)

growth media · per 1000 mL

Standard LB with salt — most common E. coli growth medium.

• Tryptone: 10 g
• Yeast Extract: 5 g
• NaCl: 10 g

SOC Medium

growth media · per 1000 mL

Rich recovery medium for transformations — higher yield than LB alone.

• Tryptone: 20 g
• Yeast Extract: 5 g
• NaCl: 0.5 g
• KCl (250 mM stock): 10 mL
• 1 M MgCl₂: 10 mL
• 1 M MgSO₄: 10 mL
• Glucose (add after autoclaving): 3.6 g

LB Agar plates

agar plates · per 1000 mL

LB agar for pouring individual Petri dishes.

• Tryptone: 10 g
• Yeast Extract: 5 g
• NaCl: 10 g
• Agar: 15 g

YPD Agar plates

agar plates · per 1000 mL

Yeast extract / peptone agar plates for yeast colony work.

• Yeast Extract: 10 g
• Peptone: 20 g
• Dextrose (add after autoclaving): 20 g
• Agar: 20 g

PBS soft agar

agar plates · per 500 mL

PBS soft agar for overlay / individual plating workflows.

• NaCl: 4 g
• KCl: 0.1 g
• Na₂HPO₄: 0.72 g
• KH₂PO₄: 0.12 g
• Agar: 3.5 g

LB Agar + Ampicillin (100 µg/mL)

selection plates · per 1000 mL

LB agar plates with 100 µg/mL ampicillin — standard bla/AmpR selection.

• Tryptone: 10 g
• Yeast Extract: 5 g
• NaCl: 10 g
• Agar: 15 g
• Ampicillin stock, 100 mg/mL (add at 50 °C): 1 mL

LB Agar + Kanamycin (50 µg/mL)

selection plates · per 1000 mL

LB agar plates with 50 µg/mL kanamycin — KanR/NeoR selection.

• Tryptone: 10 g
• Yeast Extract: 5 g
• NaCl: 10 g
• Agar: 15 g
• Kanamycin stock, 50 mg/mL (add at 50 °C): 1 mL

LB Agar + Chloramphenicol (25 µg/mL)

selection plates · per 1000 mL

LB agar plates with 25 µg/mL chloramphenicol — CmR selection.

• Tryptone: 10 g
• Yeast Extract: 5 g
• NaCl: 10 g
• Agar: 15 g
• Chloramphenicol stock, 25 mg/mL in ethanol (add at 50 °C): 1 mL