Wednesday, July 2, 2014

P. larvae germination in BAD medium assay SOP

Below is the yet tentative protocol :

Germination Determination of P. larvae in Bee Artificial Diet

This procedure is to determine if P. larvae is able to germinate in the Bee Artificial Diet (BAD) before it is fed to the larval honey bees.
  1. Perform P. larvae Spore Extraction protocol.
  2.  Perform 10 fold serial dilution of heat killed Spore Stock.
  3. Inoculate 100 uL of each dilution into 400 uL of BAD in Eppendorf tubes.
Inoculate 100 uL in 400 uL of sterile ddH2O and 400 uL of BHI broth with 1ug/ml of Thiamine as negative and positive controls, respectively.

  1. Incubate at 35C for 2 hours.
  2. Perform 10 fold serial dilution of each sample and perform drop plat protocol to quantify CFU/mL.

//EWW

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