The New Image

Caption: This images shows cultured HUVECs where the centre cell was injected with Lucifer Yellow (LY) fluorescent dye and left for 1minute. The spread of LY from the central cell to the neighbouring cells demonstrate a functional direct communication channel via gap junctions between HUVECs.
Contributed by Chiuhui Mary Wang.

The New Featured Protocol

This week’s new featured protocol comes from Professor Kenneth Peterson at the University of Kansas Medical Center. The protocol describes a method for preparing intact yeast artificial chromosome (YAC) DNA for the transgenesis of mice. Large capacity cloning vectors such as YACs are useful as they enable the entire locus and all its flanking cis-regulatory elements to be included in the construct, however, physical shearing of the DNA must be avoided during purification and microinjection in order to obtain animals bearing full-length transgenes. This method uses pulsed field gel electrophoresis to separate YAC DNA from yeast chromosomal DNA and then employs a second dimension electrophoresis to concentrate the YAC DNA. Maintenance of the YAC in an agarose gel matrix until the final steps before microinjection avoids damage to the DNA, then the matrix-embedded YAC DNA is enzymatically digested to produce a solution that can be microinjected.

Preparation of intact yeast artificial chromosome DNA for transgenesis of mice

Thank you to Hannah Clark our resident expert for supplying the text for this section.

News from the Protocols Network

My happiness index increased to almost-maximum earlier this week when the new Protocols Network submission page went live. You can check it out here:

protocols.nature.com

Happy New Year etc

Well, it is almost the end of the year, and I think that this will be the last post on my blog in 2007. I am off to Chicago for Christmas to visit with a friend and am looking forward to it despite being ever so slightly apprehensive about the coldness!

Enjoy the festivities and best wishes for 2008!