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10.B. Homogenization Method
The SEV mode has an option that allows homogenization of tissue samples prior to running the purification
method. This method must be user-validated to match the tissue sample type. In these methods, a small volume of
lysis buffer is added to an elution tube, and the tube containing the plunger is placed in the elution well. Homog-
enization occurs in this tube. Extra plungers (Cat.# AS5201) and elution tubes (Cat.# AS5101) are needed for
this method.
Guidelines for Using the Homogenization Method
Volume of Lysis Buffer: This depends on whether the sample floats in the lysis buffer. If the sample floats, use
•
200µl of lysis buffer. If the sample sinks, use 300µl of lysis buffer.
•
Sample Heating: The sample can be heated during homogenization to enhance the release of nucleic acid.
The user decides whether this is required.
•
Homogenization Time: The user must establish homogenization time. The maximum time is 10 minutes.
Running Homogenization Methods
In the Protocols screen, select Other followed by Homogenization.
Figure 42. SEV Protocols screen.
For SEV, insert the sample and lysis buffer into the elution tube, and place the elution tube into the elution rack.
Place the plunger in the tube, and press the Run/Stop button.
After the homogenization steps are complete the nucleic acid may be isolated using the appropriate purification
method. If heating was used during the homogenization method, allow the elution tube slots to cool before setting
up the purification run.
52
Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA · Toll Free in USA 800-356-9526 · 608-274-4330 · Fax 608-277-2516
TM320 · Revised 4/15
Figure 43. LEV Protocols screen.
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