cDNA was prepared using the FastLane Cell cDNA Kit, with (+RT) or without (-RT) reverse transcriptase. The cDNA was quantified using a gene expression assay for β-actin. The -RT plot in indicates that no genomic DNA was detected.
cDNA was prepared from a 48-well plate containing HeLa cells using the FastLane Kit. Expression of tubulin B (structural protein) was analyzed by real-time two-step RT-PCR.
MCF-7 cells were transfected with various CDC2 siRNAs (S1, S2, or S3) or nonsilencing siRNA (NS). After 48 hours, cDNA was prepared using the FastLane Kit. Expression of CDC2 (cell cycle protein) relative to GAPDH (endogenous control) was analyzed by real-time two-step RT-PCR. The relative expression of CDC2 in untreated cells (UT) was set to 100%.
In real-time two-step RT-PCR analysis of various genes important to cancer research in HeLa cells, lower CT values were achieved when cDNA was prepared with the FastLane Kit instead of a similar kit from Supplier AIV.