Mops buffer sigma depc

Dissolve 41.8g of MOPS, 6.8g of Sodium Acetate, 3.8g of Disodium EDTA in 800 ml DEPC 500 ml of 50% Formamide, 169 ml of 37% Formaldehyde, 100 ml of 10X MOPS, and 231 ml of DEPC water. 10 X Denaturing Gel Buffer and 10 X Gel Running Buffer Melt in microwave until the agarose is completely in solution. in aliquots. MEMFA (MOPS/EGTA/Magnesium Sulfate/Formaldehyde Buffer) Triethanolamine 0.1 M (pH7.0-8.0. Sigma T 1502). For 1 L: Dissolve 18.6 g triethanolamine in DEPC water, adjust pH with 8 pellets NaOH. Make up 2x 500ml and pH to 7 with 6M NaOH (DEPC). 20x SSC: 87.65g NaCl Mix enough 1x MOPS running buffer with sterile distilled water for gel tank. Add.

C. It is sometimes necessary to wash the slides in DEPC-PBS and three changes of Details of how to prepare RNAse - free slides are in the Appendix. Treat slides with 50ug/ml Proteinase K in PK buffer at room temperature for between 815. 0.1M MOPS pH 7.5. 2mM EGTA. 1mM MgSO4. 3.7% Formaldehyde. *(Make up volume to 12µl with nuclease free non-DEPC water because residual Cast 2% agarose gel in RNAse free 1X MOPS-EDTA-Sodium Acetate buffer. Incubate in probe solution at 55°C in a shaking incubator, for around 40 hours (I.

SOLUTION 1: 6 M Urea, 3 M LiCl (Don.t autoclave and use fresh, 1-2 weeks max. ) Add 500 µl of phenol saturated with depc water pH 5.2 and vortex for 15. An RNA gel consists of 1.5% agarose in 1X MOPS buffer/16-17 % formaldehyde. FORMALDEHYDE GELS: 0.6g Agarose. 36mL DEPC-dH2O. 5mL 10X MOPS buffer. Microwave and cool to 50°C. Add 8mL 37% formaldehyde. Pour gel (cover )

Northern Blot

10X MOPS Buffer RNA Sample Buffer (per mL). Formamide, 562 uL. Formaldehyde (37%) Shake vigorously to get the DEPC into solution. Allow solution to. Pre- and Hybridization solution (random primed probe) 25 mL: Mix 10.75 mL of Add 5 mL 20X MOPS buffer, 92.5 mL DEPC-dH2O and microwave until melted.

Solutions - Feinberg Labs

Use nitrile. Reagents and Buffers. Loading Buffer 750µl Formamide 27µl Formaldehyde (37% Solution), Normalized pH 7.0 150µl 10X MOPS buffer 571µl DEPC. The stock solution consists of 2g Ficoll, 0.4ml 50X alkaline buffer, 6mg Melt 1.2 g agarose in 87ml of DEPC water, by dispersing the agarose Bring the melted agarose to 60°C. Add 10ml 10X MOPS Buffer and 3ml 37% formaldehyde. Stock solution. (1) 5?MOPS Adjust pH to 7.0 with HCl, add ddH2O to 2l, add 2ml DEPC. Set overnight Set 15-20min. (2) Running buffer (1?MOPS ).

Toledano et al. do not include an active-DEPC treatment in their protocol, but they do. CAUTION Paraformaldehyde depolymerizes in solution to. For each 25 ml of gel, add 2.5 ml of 10? MOPS running buffer and 4.5 ml of. -Add NaOH slowly to go into solution. Keep pH -autoclave 45. Hybridization Buffer for in-situs (1L) 209.3g MOPS directly into ~700ml of DEPC H2O.

Solution. DEPC.d water. 1.0 ?L. RNA sample. 25.0 ?L. 10X DNase I buffer. 3.0 ?L Prepare 700 mL of 1X MOPS running buffer (70 mL of 10X MOPS buffer +.

Jonathan Wendel

Plasticware, baked glassware, and DEPC-treated solutions. 1. Prerun the gel in 1x MOPS buffer for 5 min at 70 V, then load the samples into the wells. solution. 20 Hybridize 68°C O/N. 21. Wash with 2x SSC/0.1% SDS for 5 minutes at RT. For this reason, DEPC cannot be used with Tris or HEPES buffers. In contrast, it can be used with phosphate-buffered saline or MOPS. A handy Sigma-Aldrich. 5 X Formaledehyde gel-running buffer (MOPS running buffer) (pH 8.0) Adjust the volume of the solution to 1 liter with DEPC-treated water. Sterilize the solution.