Selected article for: "agarose gel and initial denaturation"
Author: Klaile, Esther; Klassert, Tilman E; Scheffrahn, Inka; Müller, Mario M; Heinrich, Annina; Heyl, Kerstin A; Dienemann, Hendrik; Grünewald, Christiane; Bals, Robert; Singer, Bernhard B; Slevogt, Hortense
Title: Carcinoembryonic antigen (CEA)-related cell adhesion molecules are co-expressed in the human lung and their expression can be modulated in bronchial epithelial cells by non-typable Haemophilus influenzae, Moraxella catarrhalis, TLR3, and type I and II interferons
  • Document date: 2013_8_14
    • ID: 1fmsipqu_19
    Snippet: PCRs of the cDNAs were carried out on a S1000TM Thermal Cycler (BioRad) in a 25 μl reaction volume containing 0.2 μM primers, 1 U Taq DNA polymerase (5-Prime) and 200 μM dNTPs. Thermal conditions included an initial 95°C denaturation step for 3 min, and then 35 cycles of 10 s at 94°C, 30 s at 60°C and 30 s at 72°C. The resulting PCR products were separated on an ethidium bromide containing agarose gel and visualized under a UV-transilumina.....
    Document: PCRs of the cDNAs were carried out on a S1000TM Thermal Cycler (BioRad) in a 25 μl reaction volume containing 0.2 μM primers, 1 U Taq DNA polymerase (5-Prime) and 200 μM dNTPs. Thermal conditions included an initial 95°C denaturation step for 3 min, and then 35 cycles of 10 s at 94°C, 30 s at 60°C and 30 s at 72°C. The resulting PCR products were separated on an ethidium bromide containing agarose gel and visualized under a UV-transiluminator to confirm the expected amplicon size. To verify the identity of amplified PCR fragments of the CEACAM genes and the different splicing variants of CEACAM1, PCR-products were purified and sequenced (Eurofins MWG Operon, Ebersberg, Germany). Sequences were aligned with the corresponding NCBI-reference transcripts using ClustalX [45] .

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