Lin and groups B probes..
This is a picture of Lin's miniprep gel,
which was later used in a Southern hybridisation step.
Lin used ApE to calculate the sizes of the fragments in the Lambda BamHI + EcoRI
digest which was run in lane L.
And she uses the BamHI-EcoRI restriction map of Lambda DNA (see the Lambda DNA
page of the SCLResources site) to know which Bam probe or which Eco probe could
detect the various Lambda inserts in her clones.
To decide which probe to use to get more
information about her recombinant clones, she shows in this picture the various
possibilities.
Situations A to L show the DNA bands which give
a Southern hybridisation signal when the blot
is hybridised using the following group B probes (i.e. made with DNA
fragments purified from gel slices):
A. Lambda Bam A fragment
B. Lambda Bam B fragment
C. Lambda Bam C+D fragments
D. Lambda Bam E+F fragments
E. Lambda Eco A fragment
F. Lambda Eco B fragment
G. Lambda Eco C+D fragments
H. Lambda Eco E fragment
I. Lambda Eco F fragment
Her Lambda digest in lane L was not
heated at 70 C before loading the gel slot, so she expects
the presence of some cos-site
containing fragments (J). The cos-ends
containing fragments may join to each other by forming the cos-site and form
bigger (cos-site
containing) fragments. These ones (K) did not,
and still have their free cos-end. Others have formed the bigger
cos-site containing molecules present in this (J) band.
Note: this bigger fragment can be cloned and actually, Lin found it as an
insert in her clone 5.. (L).