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Figure 2 | Breast Cancer Research

Figure 2

From: Genetic variation in the genome-wide predicted estrogen response element-related sequences is associated with breast cancer development

Figure 2

Transcription (mRNA) of NRCAM detected by reverse transcriptase-polymerase chain reaction (RT-PCR) assay is seen in the estrogen receptor (ER)-positive breast cancer cell line MCF-7, but not in the ER-negative breast cancer cell line MDA-MB-231. GAPDH is the positive (mRNA) control of RT-PCR. For RT-PCR, total RNA was extracted from cell lines using the Arcturus PicoPur RNA Isolation Kit (Applied Biosystems, Forster City, CA, USA), and to convert RNA into cDNA, reverse transcription was performed for 70 minutes at 42°C in a reaction volume of 20 μl containing 1 μg of RNA, 10 mM random oligo(dT) primer (Promega), and 5 U of SuperScript II reverse transcriptase (Gibco-BRL, Gaithersburg, MD, USA). The presence of a cDNA band of the appropriate molecular weight was determined on 1% agarose gel after electrophoresis.

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