Archived Protocol

<BODY BACKGROUND="../gif/chalk.gif" TEXT=00009C LINK=8E236B VLINK=CC3299 ALINK=FF1CAE>  <HTML> <HEAD> <title>Stock Solution</title> <DL><DD> <H3>Stock Solution</H3> </HEAD> <H3>BSA</H3> Stock BSA from BRL comes as 50 mg/1000 µl. Add 4 ml of H2O for a final concentration of 10 mg/ml. Store at -20 C. <H3>EtBr (10 mg/ml)</H3> Dissolve 100 mg EtBr in 10 ml H2O. Stir well and store at 4 C in an amber bottle. <H3>0.5 M EDTA (pH 8.0): 500 ml</H3> Dissolve 95.05 g of Na2EDTA in appr. 400 ml H2O. Adjust pH to 8.0 with 6 N NaOH. Then bring volume to 500 ml and autoclave. <H3>0.5 M Na3EDTA: 1L</H3> Add 186.2 g (?) to 900 ml H2O. Adjust pH with conc HCl (or NaOH ?) to 8.0, and bring up to 1 L. <H3>5 M LiCl 50 mM MOPS (pH 8.0)</H3> Conc <BR> 5 M LiCl 21.2 g <BR> 50 mM MOPS 1.05 g <BR> H2O up to 100 ml <BR> Adjust pH 8.0 with NaOH. Store at 4 C. <H3>50 % PEG</H3> Add 100 g PEG (Polyethylene Glycol, appr. MW 8.000) to 50 ml H2O and stir for about one hour. Then bring up to 200 ml with H2O. <H3>3 N Perchloric acid</H3> Dilute 49.2 ml of perchloric acid up to 200 ml with H2O in a hood. Stable at room temperature for several month. Treat with care. <H3>1 N NaOH</H3> Dissove 40 g of NaOH in 1000 ml H2O. <H3>1M NaHPO4</H3> Stock <BR> NaH2PO4.H2O (monobasic monohydrate) 47.5 g <BR> Na2PO4.7H2O (dibasic 7-hydrate) 42.2 g <BR> H2O to 500 ml <H3>10 M NH4Ac</H3> Add 400 ml H2O to a 1 Kg bottle NH4Ac, dissolve, and adjust to 1300 ml. <H3>Tris 1 M pH 8.0</H3> Stock <BR> Tris-HCl 88.8 g 177 g 355.2 g <BR> Tris-Base 53.0 g 106.0 g 212.0 g <BR> H2O 1L 2 L 4 L <BR> <H3>Tris 1 M pH 7.5</H3> Stock <BR> Tris-HCl 127 g 254 g 508 g <BR> Tris-Base 23.6 g 47.2 g 94.4 g <BR> H2O 1 L 2 L 4 L <H3>76 % EtOH 10 mM NH4Ac</H3> Conc. Stock Volume <BR> 75 % 95 % EtOH 80.0 ml <BR> 0.2 M 5 M NH4OAc 0.2 ml <BR> H2O 19.8 ml <BR> 5 M NH4Ac stock = 38.55 g/100 ml <H3>76 % EtOH 0.2 M NaAc</H3> Conc. Stock Volume <BR> 76 % 95 % EtOH 160 ml <BR> 0.2 M 3 M NaAc 13.33 <BR> H2O 26.67 <BR> 3 M NaOAc stock = 40.83 g/100 ml <H3>0.2 N NaOH 0.6 M NaCl</H3> 1L 2L 3L 4L <BR> NaOH 8.0 g 16.0 g 24.0 g 32.0 g <BR> NaCl 35.04 g 70.08 g 105.2 g 140.2 g <H3>0.5 M Tris (pH 7.5) 1.5 M NaCl</H3> 1L 2L 3L <BR> Tris-HCl 63.5 g 127.0 g 190.0 g <BR> Tris-Base 11.8 g 23.6 g 35.4 g <BR> NaCl 87.6 g 175.0 g 262.8 g <H3>25 mM NaHPO4</H3> 1M NaHPO4 100 ml <BR> H2O to 4 L <H3>10 X Treacking Dye</H3> Conc. <BR> 50 mM Tris pH 8.0 <BR> 50 mM Na3EDTA <BR> 25 % Glycerol <BR> 5 % Ficoll <BR> 0.1 % Xylenol Cyanol <BR> 0.1 % Bromophenol Blue <BR> 1 % SDS <H3>Glycerol Solution </H3> Glycerol 650 ml <BR> 1 M MgSO4 100 ml <BR> 1 M Tris (pH 8) 25 ml <BR> H2O up to 1 L <BR> Mix well and autoclave for 15 min at 121 C. <H3>0.1 M Spermidine Solution</H3> Dissolve 255 mg spermidine (Sigma #S2501) in H2O to a final volume of 10 ml. Store at -20 C. <H3>Saline-MOPS</H3> 1X 1L 1X <BR> 0.85 % NaCl 8.5 g <BR> 50 mM MOPS 6.3 g MOPS free acid <BR> 4.7 g MOPS Na salt <BR> pH 7 autoclave and store at room temperature. <H3>Proteinase K - 10 mg/ml in TE</H3> Weigh out 10 mg proteinase K (Boeringer Manheim #161 497) and dissolve in 1 ml TE. The solution can be used immediately or aliquoted and stored at -20 C <H3>RNase A and RNase T1 </H3> Dissolve 100 mg RNase A (bovine pancrease: Sigma #R4875), if desired. add 5,000 units of RNase T1 together, in 10 ml of 10 mM Tris 15 mM NaCl. Boil for 15 min and allow to cool slowly to room temperature. Distribute 1 ml aliquote into 1.5 ml MFT, and store at -20 C. If </DL> </BODY> </HTML>