Selected article for: "High fidelity dna polymerase and supplementary table"
Author: Tromas, Nicolas; Zwart, Mark P.; Forment, Javier; Elena, Santiago F.
Title: Shrinkage of Genome Size in a Plant RNA Virus upon Transfer of an Essential Viral Gene into the Host Genome
  • Document date: 2014_2_20
    • ID: 5fejitls_15
    Snippet: Subsequently, the full genome was reverse transcribed using M-MLV RT (Fermentas) and primers 97-101-R, 67-77-R, and 40-45-R and PCR amplified using the high-fidelity DNA polymerase Phusion (Finnzymes) and three pairs of primers 2-10-F/40-45-R, 45-48-F/67-77-R, and 73-80-F/97-101-R (supplementary table S1, Supplementary Material online). By using these pairs, we ensured that the mRNA from the transgene was not amplified and that we obtained only T.....
    Document: Subsequently, the full genome was reverse transcribed using M-MLV RT (Fermentas) and primers 97-101-R, 67-77-R, and 40-45-R and PCR amplified using the high-fidelity DNA polymerase Phusion (Finnzymes) and three pairs of primers 2-10-F/40-45-R, 45-48-F/67-77-R, and 73-80-F/97-101-R (supplementary table S1, Supplementary Material online). By using these pairs, we ensured that the mRNA from the transgene was not amplified and that we obtained only TEV sequences from viral genomes. PCR products were resolved on 1% agarose gels to verify the presence of deletions. For both genotypes, TEV and TEV-ÁNIb, the three amplicons were purified and sequenced by GenoScreen (Lille, France) using BIGDYE 3.1 and a 96-capillars ABI3730XL sequencing system (Applied Biosystems) with overlapping readouts using the same set of primers as in Agudelo-Romero et al. (2008) . Contigs were assembled using GENEIOUS version 4.8.

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