Reagent Prepration Lab Report

Reagent prepration:

0.7M NaCl

Add 14ml of 5M NaCl to 86ml of distilled water. Sterilise by autoclaving.

CTAB/NaC1 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 NaC1 solution.

PCI (phenol:chlorophora:isoamyl alcohol) ---------100ml

This is a solution of phenol, chloroform, and isoamyl alcohol, in a ratio of

25:24:1; with a pH of 7.5-8.0

• Proteinase K (20mg/mL)

Dissove in water and store at -200C.

Store Proteinase K at -20°C.

• 10% SDS --------------------------- 100ml

Dissolve 10 g of SDS (Sodium Dodecyl Sulfate) in 80 mL of H2O,

Heat to 680C to dissolve & adjust pH to 7.2 by adding few drops of conc. HCl and adjust the volume to 100 mL with H2O.

• 5 M sodium chloride -----------------100ml

Dissolve 29.22g of sodium chloride (NaC1; M.W. 58.44) in 80 ml of H2O.

Adjust the volume to 100 ml with H2O.

Sterilize by autoclaving.

0.5 M EDTA pH 8.0 (ethylenediaminetetraacetic acid)------------------100ml

Add 16.81 g of disodium ethy1enediaminetetraacetate to 80 mL of H2O. Stir vigorously on a magnetic stirrer. Adjust the pH to 8.0 with NaOH (approximately 2 g of NaOH pellets). Adjust the volume to 100 ml with H2O.

NOTE: The disodium salt of EDTA will not go into solution until the pH is adjusted to approximately 8.0.

Sterilize by autoclaving.

1M Tris-----------------------------------100ml

Dissolve 12.11 g of Tris base in 80 ml of H2O. Adjust pH to 8 by adding 4.2ml concentrated