leukemic cells will be analyzed for vector presence 
and the number of leukemic clones containing 
vector. 
6 . 21 ) 
Studies to be performed 
6.211) Pre-harvest samples (<3 weeks 
prior to harvest) . 
а. Bone marrow aspirate and 
biopsy 
light microscopy 
cytogenetics 
6.212) Pretreatment - Baseline studies 
to test for the presence of 
retroviral vector and 
retrovirus in the untreated 
bone marrow cells of the 
patient (PCR for neo R and viral 
env gene, and 3T3 
amplification) and serum 
antibodies to viral proteins 
(Western blot) (see Section 
б. 3). 
a. Bone marrow cells at 
harvest (untreated) 
10 8 cells in preservative- 
free heparin: 
2xl0 6 cells for neo R PCR 
2xl0 6 cells for viral env 
PCR 
2xl0 6 cells for 3T3 
amplification 
lxlO 7 cells for Southern 
blot analysis 
2 x 1 0 6 cells for 
methylcellulose assay 
remainder - store in 
liquid nitrogen 
b. Blood (day of harvest) 
10 ml red top tube, store 
serum at -70 °C run 
Western blot after d 100 
Recombinant DNA Research, Volume 15 
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