Molecular cloning : a laboratory manual by Joseph Sambrook; E F Fritsch; Tom Maniatis

By Joseph Sambrook; E F Fritsch; Tom Maniatis

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0) immediately before use. 0 before dissolving the protein. 0. Media Containing Agar or Agarose Prepare liquid media according to the recipes given. 05 kg/cm 2) on liquid cycle. When the medium is removed from the autoclave, swirl it gently to distribute the melted agar or agarose evenly throughout the solution. Be careful! The fluid may be superheated and may boil over when swirled. , antibiotics). To avoid producing air bubbles, mix the medium by swirling. Plates can then be poured directly from the flask; allow approx.

Centrifuge the tube at maximum speed for 30 seconds at 4°C in a microfuge. Store the unused portion of the culture at 4°C. Remove the medium by gentle aspiration, leaving the bacterial pellet as dry as possible. Resuspend the bacterial pellet in 350 µl of STET. Add 25 µl of a freshly prepared solution of lysozyme. Close the top of the tube and mix the contents by gently vortexing for 3 seconds. Place the tube in a boiling water bath for exactly 40 seconds. 2003 22:34:41] Molecular Cloning | Protocol 4Print Version 7.

1 g of Tris base in 800 ml of H2O. Adjust the pH to the desired value by adding concentrated HCl. 0 42 ml (1 M) Allow the solution to cool to room temperature before making final adjustments to the pH. Adjust the volume of the solution to 1 liter with H2O. Dispense into aliquots and sterilize by autoclaving. If the 1 M solution has a yellow color, discard it and obtain Tris of better quality. The pH of Tris solutions is temperature-dependent and decreases approx. 03 pH units for each 1°C increase in temperature.

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