We currently have several problems related to transforming Agrobacterium
for complementation. First we have a rather large region which we would
like to transform which is not represented in at least two cosmid
libraries. We have therefore been trying to subclone lambda clones (size
16-20 kbp) from the region into transformation vectors. Has anyone else had
to go this route? Unfortunately we don't yet have the ideal vectors for
this. Does anyone have a transformation vector with an SfiI site in it?
Or better yet does anyone have a cosmid vector with an SfiI site that will
accept and package 16kbp inserts? We currently have pOCA18 and pCIT30,
both of which are not ideal for our purposes.
Thanks for help with any of these questions,