Prepare solutions for 10 mM Ce(IV) positive control reaction Prepare each soluti
ID: 483606 • Letter: P
Question
Prepare solutions for 10 mM Ce(IV) positive control reaction Prepare each solution in 1 mL HEPES. The final volume will be 5 mL, so masses are shown for a 5 mL solution. Store separately in centrifuge tubes until ready to start reaction. To start a reaction, combine the three individual solutions and dilute to 5 mL with HEPES. Ce IV) @ 10 mM = 27.41 mg (for 5 ml) dTpdA @ 1 mM = 2.85 mg (for 5 mL) NaCl @ 100 mM = 29.22 mg (for 5 mL) Prepare solutions for 10 mM UA-Asc experimental reaction Prepare each solution in 1 mL Tris-HCI. The final volume will be 5 mL, so masses are shown for a 5 mL solution. Store solutions separately in centrifuge tubes ready to start reaction. To start a reaction, combine the four individual solutions and dilute to 5 mL with Tris-HCl. dTpdA @ 1 mM = 2.85 (for 5 mL) NaCl @ 100 mM = 29.22 mg (for 5 mL) UA @ 10 mM = 21.21 mg (for 5 mL) Asca @ 10 mM = 8.81 mg (add Chelex to soln; shake for 10 min: microfilter into new tube) (for 5 mL)Explanation / Answer
Ans 1 Preparation of HEPES buffer
This buffer is used in the positive control reaction. To prepare the buffer we will add 2.38g of HEPES to approximately 425 ml of water in a 2l flask. Add a few drops of chelex resin to the solution and stir for few hours.Aftwerwars filter the solution and adjust the final pH to 6.9 to 7.1. At last bring the volume to 500ml with water.
Ans 2 Preparation of Tris HCl buffer
THis buffer is used in the experimental reaction.To prepare the buffer we will add 1.2114 g of Tris HCl to approximately 425ml of water in a 2L flask .Add few drops of chelex resin and stir the solution for few hours.Finally filter the solution and adjust final pH to 7.9 to 8.1 .Brin the volume to 500 ml with water.