Preparation of buffer
Materials required
Buffer preparation (for 1 litre diluent)
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Take the triple distilled water in a sterile beaker according to the requirement
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Weigh the chemicals accurately
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Add the chemicals one by one and dissolve it by magnetic stirrer by required speed
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Finally add the glycerol at 7% level ie. 70 ml
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Completely mix the glycerol by magnetic stirrer for at least 20 minutes
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Care should be taken while stirring that air bubbles should not be formed by over speed of the stirrer
Sterilization of buffer
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The prepared buffer is sterilized by autoclaving at 100 ⁰ C for 15 min at 5 psi.
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Cool the buffer to room temperature
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Generally the buffer is prepared on previous day evening, sterilized and kept under laminar air flow over night.
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The diluent is prepared by adding egg yolk in the morning immediately before semen collection.
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Last modified: Monday, 4 June 2012, 10:15 AM