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s. HeLa cells were lysed at a concentration of 1000 cells/l using the Cells-to-Signal Lysis Buffer. 100 l aliquots of the lysate were stored for two months at -80C or -20C. Ambion's RNAqueous Kit was then used to isolate total RNA from each aliquot along with an aliquot of lysate that was freshly prepared (not stored). RNA isolated from an equivalent number of cells processed using the RNAqueous procedure is shown on the far right. The RNA samples were analyzed on an RNA LabChip using the Agilent 2100 bioanalyzer and show rRNA bands.


Complete Kit for Cell Lysis and cDNA Synthesis
The Cells-to-Signal Kits contain reagents for 30 or 100 reverse transcription reactions, including M-MLV RT, oligo(dT)18 primers, and random decamer primers. Up to 5 x 104 cells per reaction can be used for the lysis step; up to 30% of the RT reaction volume can consist of cell lysate. Ambion scientists have used the Cells-to-Signal Kit with cell lines including HeLa, HeLa S3, 293, A549, MCF-7, K562, SKNAS and NHDF-neo cells, and we continue to test other cell types. The kit contains a positive control RNA and complementary primers for optimizing the procedure. This control can be added with the lysis solution to monitor the efficiency of the procedure. SuperTaq Polymerase is available separately.


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