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. Author manuscript; available in PMC: 2013 Oct 31.
Published in final edited form as: Methods Mol Biol. 2008;461:10.1007/978-1-60327-483-8_31. doi: 10.1007/978-1-60327-483-8_31

Fig. 4.

Fig. 4

Method for backfilling nuclei into a transplantation needle. (A) The diluted reaction mix is drawn into a clipped yellow tip containing about 1cm of Tygon tubing. (B) The yellow tip containing the Tygon tubing and dilute sperm nuclei is carefully detached from the pipetman and connected to the back of needle using the tubing. The needle is gently loaded with the dilute sperm nuclear reaction by gravity. This is done by slowly increasing the angle of the yellow tip/tubing/needle so that the mixture flows gently into the needle. Once the needle is complete filled, the needle is detached from the yellow tip and the needle is ready to connect to the infusion pump. The remaining sperm mixture can be set aside horizontally and used to reload another needle, if two people are injecting simultaneously or if a needle is accidentally damaged or blocked.