Reply to: RE>bluescript
I don't know the "answer", but I suggest running a few controls -
[I assume you are using purified insert for your heterologous probe, nonethe
(i) make southern blots of your putative "clones" that contain lanes of the
digested original LAMBDA and the released PLASMID
(ii) label your probe's vector, and screen the southern blots with that as well
(iii) purify/isolate a few "negative" (non-hybridizing) plaques, which contain
an insert, and include these in the above southern blots -- along with
"wildtype" lambda zap and pBS -- for screening with your heterologous probe
(and its purified vector).
(iv) use another probe, guaranteed to find a clone (e.g., ssRUBPCase, tubulin,
actin), that is in the same vector as your heterologous probe. Then screen
your old filters, southern blots, etc.
(v) screen a genomic southern blot with your heterologous probe to see if any
homology exists [this will aslo tell you about # of fragments, size(s), best
enzme to use for cloning, etc.]. Again, I assume this has already been done.
If not, consider doing this first.
** if an answer comes in, and you can avoid any or all of the above, please let
me know. Good luck.
Vance_Baird at Quickmail.Clemson.EDU