protocols. (Section 9.0 Multiple Myeloma and CML protocols. 
Section 11.0 Breast Cancer protocol) 
6.3 20 cc of blood will be drawn weekly before engraftment, 
monthly after engraftment, and at the time of relapse 
as described in the original protocols for FACS 
analysis, clonagenic assays and PCR analysis for tumor- 
specific markers. Marrow will be obtained for these 
purposes every one-two weeks prior to engraftment, 
every one-three months after engraftment, and at the 
time of relapse, also as described in the protocols. In 
patients entered onto the genetic marking amendment, - 
these samples will also be analyzed using PCR and/or 
Southern blotting for the transduced Neo gene and for 
wild-type helper virus. 
Bone marrow and blood samples will be plated in 
methylcellulose , and individual and pooled 
hematopoietic colonies of all lineages will be plucked 
and analyzed for the presence of the transduced Neo 
gene via PCR. The percentage of transduced progenitors 
will also be assessed by growth of Neo-resistant 
colonies in methylcellulose culture. Bone marrow and 
peripheral blood cells will also be sorted via FACS 
into T-cell, B-cell and specific myeloid populations, 
and analyzed via PCR for the transduced gene (or 
Southern blotting if transduction efficiency has been 
greater than 10% in any population of cells). Insertion 
site analysis will be used to investigate clonality and 
lineage relationships if Southern blotting is carried 
out . 
6.4 CML Patients : RNA and DNA from individual colonies or 
FACS-sorted cell populations in CML patients will be 
concurrently analyzed for the tumor-specific bcr-abl 
rearrangement and the Neo gene via reverse 
transcription and PCR. 
6.5 Multiple Myeloma Patients : DNA from peripheral blood, 
bone marrow, and cell populations sorted by FACS for B- 
cell and plasma cell antigens will be analyzed for 
tumor-specific immunoglobulin gene rearrangements via 
allele-specific PCR and concurrently for the transduced 
Neo gene. 
6.6 Breast Cancer Patients : As described in section 11.13 
of the original protocol, biopsies of accessible tumor 
(skin, lymph nodes, pleural effusions, bone marrow, 
liver or lung lesions amenable to CT-guided FNA) will 
be done at the time of relapse and DNA obtained for PCR 
and Southern blot analysis of the Neo gene. As 
described in section 11.6 of the breast cancer 
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Recombinant DNA Research, Volume 16 
