June 2, 1972

Dr. Lloyd J. Old
Sloan-Keltering Inrtitute
410 lbrt 68th Street
New York City, New York

10021

Dear Dr, Old:

Bob Nowinski called today to tell me of your kind offer to collaborate with
ur in rtudier of base requence homologies between nucleic acid8 from human
breart tupora and froan Mu W.
moruring rather precisely the amount of EiuWrV-specific RNA in a variety of
amuse tlssuer, in an effort to describe the degree of tranrcriptional rea-
ulation of the large amount of MWEV DNA found in both OB and C5XBl strains
(see enclored preprint).  The umure studies indicated that we could anneal
all of a ringle stranded BNA probe -de by EfUHfv polymerarcs to RNA from
mnmry tumors, vlrur-producing manawry glands, and lactating mammary glands
from C57 mice.
rpecifie DUA nuclease and in Cat304 gradients. ScaPplete annealing war
obt8ined with spleen RNA.  About .5% of WIJA in the virus-producing tissues
le M~-rpecific, about .01% in the CS?L,MG ir MuWf'V-rpecific, and less
than .001% in the spleenr in viral-rpecific.

In view of the high stenritivity stringency of our hybridi~ation and assay
procedures, we decided tu try t@ confirm Spiegelmanfr rerultr with human
breast tirsue.  To our frank surprire,  the flirt breart tumor rent to ur
by Murul Sarbr contained 0.01% MuHm-like %NA.
100% of our single-stranded DNA probe from MyMTV to thir lINA a11 teated with
both nuclease and Csw aasays.
properties of this hybrid and find that it melt8 sharply with o Ta 4OC below
the Tm for the prerulurbly completely specific hybrid formed between the ram8
probe and BNA from a vlrus-producing mouse tumor,
in bere sequence between the probe and the 8wA it is detecting are about 6%.

A8 Bob prebably told you, VB have been

Annealing war damnatrated by army +ith a ringle-strand-

Gd

We have been able to anneal

In addition, we have rtudied the melting

This suggerts the differences

We have teated several other breart rpeclmens; all nom1 breart8 end benign
tumors have been negative, but the 4 tunor8 we have subraguently tested have
also been negative or questionable.
that the amount of tumer tissue in these 8pecLswrnr tendr to be small; con-
sequeuatly we get little IUUA (which my be contaminated by ENA from adjacent
nom1 cells) and cannot purh the level of detection to the llmitr the assay
itself can bear,  Therefore we asked Bob to approach you .bout supplying us

The problem at this point 11 principally

Dr. Lloyd J. Old
June 2, 1972
Page 2

with tlglpOrs (metastatic or primary).
eotablished human breast tuwrr lines is a very appealing one.

P will call you nert week to discuss the specifics of the amounts of tissuelr
or cells we would need.
give you same details about the technique8 involved.  (I should nwntion that
we are about to examine human tissues for MuMTV-like DESA and would want to
do the same with any tissue# you mfght provide UI).
in lew York the second week in August and perhaps could get together with
you at that time.

The sumestion that we examine your

I am enclosing SOPWI recent paperr from this lab to


               In addition, I will be

Yours truly,

?tarold E. Vanuus, M.D.
Department of Microbiology

B%i:bb

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