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The RNA was converted to cDNA using the following conditions: 37°C for 60 min, 95°C for 5 min, and 4°C hold until further processing or storage.
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According to standard CHTN procedures, samples of tumor and of healthy adjacent tissues were obtained, with a portion examined by a pathologist for diagnosis and the remaining material snap-frozen in liquid nitrogen and held at −70°C until shipment on dry ice to ASU where they were held at −80°C until further processing.
The resulting lysates were stored at −80°C until further processing.
For in situ hybridization, ovarian tissue was sampled from an ovulated female, fixed in Dietrick's fixative (10% Formaldehyde, 28. 5% ethanol, 2% glacial acetic acid) at 4°C overnight, rinsed in tap water for 1 hour and held in 50% ethanol until further processing.
Oocytes, freed of cumulus cells, were washed through 3 volumes (∼2 ml each) of TALP-Hepes, and then held in a smaller drop (∼500 µl) of TALP-Hepes covered in sterile mineral oil until further processing.
DNA samples were stored at −80 °C until further processing.
Purified RNA samples were stored at −80°C until further processing.
RNA samples were stored at −80 °C until further processing.
The filters were immediately frozen at −80 °C until further processing.
Filtrate was then stored in −20°C until further processing.
Samples were stored at -20°C until further processing.
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