Luckau Protocols:Tris-Cl

Tris-Cl Protocol

Tara K. Luckau's Home Page Conservation Genetics Lab Notebook Tara's Protocols

Purpose

• Tris-Cl is a buffering agent (made from Tris base and HCl)
• We use it as a stock solution for Low TE

Info

• tris [hydroxymethyl] amino methane (C₄H₁₁NO₃, 2-Amino-2-hydroxymethyl-propane-1,3-diol)
• MW = 157.56 g/mol (MW = molecular weight, same as formula weight, FW)

Protocol

Calculations

• [math]\displaystyle{ 500 \mathrm{mL} \bullet \tfrac{0.5 \mathrm{mol}}{\mathrm{L}}\ \bullet \tfrac{\mathrm{L}}{1000 \mathrm{mL}}\ \bullet \tfrac{157.56 \mathrm{g}}{\mathrm{mol}}\ = 39.39 \mathrm{g} }[/math]

• [math]\displaystyle{ 500 \mathrm{mL} \bullet \tfrac{1 \mathrm{mol}}{\mathrm{L}}\ \bullet \tfrac{\mathrm{L}}{1000 \mathrm{mL}}\ \bullet \tfrac{157.56 \mathrm{g}}{\mathrm{mol}}\ = 78.78 \mathrm{g} }[/math]

To make [math]\displaystyle{ \tfrac{1}{2} }[/math] liter of 0.5M or 1M Tris-Cl, pH 8.0

Concentration	Tris-Cl
0.5M	39.39 g
1M	78.78 g

1. In 1 L beaker, mix Tris-Cl in 400 mL H₂O
2. Put beaker on hotplate with stirbar until dissolved
3. Using concentrated aqueous HCl to adjust to pH 8.0
4. Add H₂O to bring volume to 500 mL
5. Autoclave solution, store at room temperature