========================== Shared genomics buffers ========================== The following buffers are used widely across genomics protocols and have practically infinite shelf life (all salts). All of these should be **0.22µm filter-sterilized** (use syringe filters) and stored near the genomics hood in 66-219. .. note:: We typically mark filtered buffers with a red Sharpie line and the word "filtered". - 50 mL **1M Tris (pH 7.5)** ======================= ================ ======== ==================== Component Concentration g / L Amount / 50 mL ======================= ================ ======== ==================== Tris-HCl 1 M 157.64 7.88 g Elga water to 40 mL NaOH to pH 7.5 ~700 uL of 12 N NaOH Elga water to 50 mL Filter sterilize ======================= ================ ======== ==================== - 50 mL **2M Tris (pH 7.5)** ======================= ================ ======== ==================== Component Concentration g / L Amount / 50 mL ======================= ================ ======== ==================== Tris-HCl 2 M 157.64 15.76 g Elga water to 40 mL NaOH to pH 7.5 Elga water to 50 mL Filter sterilize ======================= ================ ======== ==================== - 50 mL **1M Tris (pH 8.0)** ======================= ================ ======== ==================== Component Concentration g / L Amount / 50 mL ======================= ================ ======== ==================== Tris-HCl 1 M 157.64 7.88 g Elga water to 40 mL NaOH to pH 8.0 ~900 uL of 12 N NaOH Elga water to 50 mL Filter sterilize ======================= ================ ======== ==================== - 50 mL of **1M HEPES pH 7.5 stock solution** ======================= ================ ======== ==================== Component Concentration g / L Amount / 50 mL ======================= ================ ======== ==================== HEPES 1 M 238.3 11.915 g Elga water to 40 mL NaOH to pH 7.5 ~1.2 mL of 12 N NaOH Elga water to 50 mL Filter sterilize ======================= ================ ======== ==================== - 50 mL of **500 mM EDTA (pH 8.0) stock solution** ======================= ==================== ======== ========================== Component Concentration g / L Amount / 50 mL ======================= ==================== ======== ========================== EDTA 500 mM 186.1 9.305 g Elga water to ~30 mL NaOH to pH 8 ~3.75 mL of 12N NaOH Elga water to 50 mL Filter sterilize ======================= ==================== ======== ========================== The EDTA makes the solution very acidic. Adding the NaOH will help it dissolve. It will still take a lot of vortexing to get the final solution! - 50 mL of **10x TE (pH 8.0)**: ======================= ==================== ========================== Component Concentration Amount / 50 mL ======================= ==================== ========================== 2M Tris 100 mM 2.5 mL 500 mM EDTA 10 mM 1.0 mL Elga water to ~40 mL 36.5 mL NaOH to pH 8 Elga water to 50 mL Filter sterilize ======================= ==================== ========================== - 50 mL of **5M NaCl stock solution**: ======================= ================ ======== ==================== Component Concentration g / L Amount / 50 mL ======================= ================ ======== ==================== NaCl 5 M 292.21 14.611 g Elga water to 50 mL Filter sterilize ======================= ================ ======== ==================== - 50 mL of **2.5M CaCl2**: ======================= ================ ======== ==================== Component Concentration g / L Amount / 50 mL ======================= ================ ======== ==================== CaCl2 dihydrate 2.5 M 147.02 18.38 g Elga water to 50 mL Filter sterilize ======================= ================ ======== ==================== - 50 mL of **1.0M MgCl2**: ======================= ================ ======== ==================== Component Concentration g / L Amount / 50 mL ======================= ================ ======== ==================== MgCl2 1.0M 95.21 4.76 g Elga water to 10 mL Filter sterilize ======================= ================ ======== ==================== - 50 mL of **10% SDS**: ======================= ================ ==================== Component Concentration Amount / 50 mL ======================= ================ ==================== SDS 10% (w/v) 5 g Elga water to 50 mL ======================= ================ ====================