Selected article for: "agarose gel and PCR product"
Author: Abdel-Moneim, Ahmed S; El-Kady, Magdy F; Ladman, Brian S; Gelb, Jack
Title: S1 gene sequence analysis of a nephropathogenic strain of avian infectious bronchitis virus in Egypt
  • Document date: 2006_9_20
    • ID: 1in7m56w_32
    Snippet: A Dot-ELISA was performed according to [39] . Briefly, NCM of convenient size was cut, marked with waterproof ink for identification and then soaked for 10 min. in distilled water. NCM was laid on absorbent paper and airdried for 5 min. Approximately 2 μl of the extracted viral RNA was used to synthesize cDNA. Amplification of the S1 gene was performed using S1OLIGO3' (5'-CATAACTAACATAAG-GGCAA-3') and S1OLIGO5' (5'-TGAAACTGAACAAAAGAC-3') primers.....
    Document: A Dot-ELISA was performed according to [39] . Briefly, NCM of convenient size was cut, marked with waterproof ink for identification and then soaked for 10 min. in distilled water. NCM was laid on absorbent paper and airdried for 5 min. Approximately 2 μl of the extracted viral RNA was used to synthesize cDNA. Amplification of the S1 gene was performed using S1OLIGO3' (5'-CATAACTAACATAAG-GGCAA-3') and S1OLIGO5' (5'-TGAAACTGAACAAAAGAC-3') primers [10, 37] . PCR of S1 gene was performed as described [11] with the exception that extension was performed at 60°C. S1 PCR product was cut from 1.8% agarose gels, purified with the QIAquick gel extraction kit (Qiagen, Inc.) and the DNA was quantitated as described [11] . Purified RT-PCR product was sequenced in the forward and reverse directions using the same primers. Sequencing was performed as described [11] .

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