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Reagent Lab Preparation For DNA Isolation

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Reagent for DNA Isolation

  • CTAB/NaCl solution (10% CTAB, 0.7 M NaC1)

Dissolve 10 g of CTAB in 80 mL of 0.7 M NaCl solution. Stir on a hot magnetic  stirrer until the CTAB has dissolved. Adjust volume to 100 mL with 0.7 M NaCl solution.

  • 10% SDS solution

Dissolve 10 g of SDS (Sodium Dodecyl Sulfate) in 80 mL of H2O, and adjust the volume to 100 mL with H2 O.

  • 50x TAE (Tris-Acetate Buffer)

Dissolve 242 g of Tris base, 57.1 mL of glacial acetic acid, and 100 mL of 0.5 M EDTA (pH 8.0) in H2O up to 1 liter. The 50x TAE is the concentrated stock solution. Use lx TAE as working solution 0.04 M Tris-acetate, 0.001 M EDTA).

  • Loading dye  6x  (0,25%  bromophenol  blue;  0,25%  xylene  cyalol;  15%  ficoll  tipe 4000; EDTA 120 mM)
  • TE Buffer Solution (pH 7.4, 7.6, 8.0)

10 mM Tris-HCl, pH 7.4

1 mM EDTA, pH 8.0

10 mM Tris-HC1, pH 7.6

1 mM EDTA, pH 8.0

10 mM Tris-HCl, pH 8.0

1 mM EDTA, pH 8.0

  • 10 mM Tris-HC1 (pH 7.4, 7.6 and 8.0)

Dissolve 121.1 g of Tris base in 800 ml of H2O. Adjust pH to the desired value by adding concentrated HCl.

pH 7.4 add 70 ml

pH 7.6 add 60 ml

pH 8.0 add 42 ml.

Other pHs desired can be obtained by titrating the HCl. Adjust the final volume of  the solution to 1000 ml with H2O.

  • Ethidium bromide solution (10 mg/mL)

Add 200 mg of ethidium bromide to 20 mL of H. Stir on a magnetic stirrer for several hours to ensure that the dye has dissolved. Store in a light-proof container (e.g., in a falcon tube wrapped in aluminum foil) at room-temperature.

  • 5 M sodium chloride

Dissolve 292.2 g of sodium chloride (NaCl; M.W. 58.44) in 800 ml of H2O. Adjust the volume to 1 liter with H2O. Sterilize by autoclaving.

Reference:

Methods and Tools in Biosciences and Medicine. Techniques in molecular systematics and evolution, ed. by Rob DeSalle et al.

© 2002 Birkhäuser Verlag Basel/Switzerland

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