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How to make a spore syringe

A pictorial by Una

This document can be found on our website, but i hadn?t posted it here yet.

Making a spore syringe

although the photographs show the procedure in the open air we make all spore syringes in a flow cabinet!

To make a spore syringe you will need the following items:

*Inoculation loop/scalpel (something to pick up the spores)
*Spore print
*Syringe and needle (sterile)
*Small amount of sterilized water in a bottle (capped with tinfoil)
*Clean flame source (gas- or alcohol burner)
*Latex gloves (optional)

The most important thing in the whole process is to work clean. If possible perform the following procedure in a glove box or HEPA filtered environment. If you do not have access to one of these you can perform it in the open air, but chances of success are smaller. If available wear gloves that have been disinfected with alcohol 70% or lysol.
CAUTION: Most disinfectants are flammable, use extreme caution when using these in combination with an open flame!

Before you start clean the working surface with alcohol or lysol. Allow it to dry before proceeding!

The inoculation loop or scalpel is flamed until red hot, and left to cool. Alternatively you can use a resterilizes inoculation loop.

Peal back the tinfoil of the flask so that it can be removed easily but leave it covering the flask. Open the baggie with the spore print and scrape some of the spores of the paper. If the loop is too hot the spores will be killed or the plastic will melt to it.:

Deposit the spores into the water and quickly replace the tinfoil. Repeat if not enough spores have been transferred. If the spores are visible in the water there should be enough.

The syringe is assembled. Most syringes can be reused by sterilizing them in a pressure cooker. Just wrap them in tinfoil and sterilize for 25-30 minutes.

Some spore suspension is sucked up in the syringe. Pull the plunger back and forth a few times to disperse the spores in the water. Syringes can then be filled with spore suspension.

The needle cap is replaced and the syringes are put in the fridge until ready to use. Don?t forget to lable them with strain and date. Syringes can be kept like this for several months after which they become noticeably less viable.

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