This technique appeared as an article in Biotechniques, Vol.14, No.3, pp412-414. This article is a report by Kristian Gundersen, Theresa Hanley and John Merlie, on the use of micropipette bevelling to increase transgenic (pronuclear injection) embryo yield. A slurry of grit 120 silicon carbide is used in a beaker on a stir plate to provide a gentle yet abrasive force. Micropipettes drawn out previously on a vertical or horizontal puller can be further modified by the immersion of their tips into the rotating slurry at approximately 45 degrees for 15-30 seconds. This imparts an aperture which is less prone to clogging and also reduces the shearing force applies to the DNA as it is expelled through the glass needle. These factors together serve to increase the viability of injected eggs and the subsequent successful transgene integration.
The vendor for the silicon carbide grit is as follows:
Buehler Ltd
41 Waukegan Rd.
Lake Bluff, IL 60044
Toll-free Number: N/A
Phone Number: 708 295-6500
Fax Number: 708 295-7979
There is a minimum order of $50.
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