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- Fluorescein-12-dUTP (Roche Molecular Biochemical), 1 mM. Store at
-20°C.
- Formamide (Fisher). Store at 4°C.
- L-Glutamine (Sigma). Store at -20°C.
- Glycogen (Roche Molecular Biochemical), 20 mg/ml. Store at -20°C.
- Heparin (Life Technologies).
- Human COTl™ DNA (Life Technologies), 1 mg/ml. Store at -20°C.
- Low melting point agarose (Bio Rad). Store at room temperature.
- Methotrexate (Sigma). 104
M in Hank's BSS. Frozen in aliquot. Dilute
to 105
M with Hanks BSS before use.
- Oligonucleotides used in in vitro DNA amplifications. Store at -20°C
- JUNl: 5-CCCAAGCTTGCATGCGAATTCNNNNCAGG -3
- JUNI5: 5- CCCAAGCTTGCATGCGAATTC -3
- Penicillin/streptomycin (Life Technologies). Store at -20°C.
- Pepsin (Sigma), 10% stock solution (100 mg/ml) in sterile water. Store
at -20°C.
- Phytohemagglutinin (Life Technologies), 10 mg/ml. Store at 4°C.
- Pronase (Sigma), 0.5% in FHM (see below). Store at -80°C.
- Proteinase K (Roche Molecular Biochemical): 20 mg/ml in 10 mM
TrisHCI, pH 7.5. Store at -20°C.
- Random-Priming Kit: BioPrime kit (Life Technologies). Store at -20°C.
- RNase (Roche Molecular Biochemical), DNase-free: 20 mg/ml in sterile
water, boil at 100°C for 10 min. Aliquot and store at -20°C.
- RPMI (Life Technologies). Store at 4°C.
- Salmon sperm DNA (3-5), 20 mg/mL Store at -20°C.
- SKY kit (Applied Spectral Imaging, Inc.). Store at 4°C.
- T7 Sequenase II and Sequenase reaction buffer (Amersham Life
Science Inc.). Store at -20°C.
- Thermus aquaticus (Taq) DNA polymerase (Perkin Elmer), 5 VlIlL
Store at -20°C.