2008
DOI: 10.1016/j.jviromet.2007.12.014
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A protocol for the rapid isolation of full geminivirus genomes from dried plant tissue

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Cited by 117 publications
(58 citation statements)
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“…Viral DNA was isolated from symptomatic leaf material at between 20 and 127 days (median, 47 days) postinoculation (p.i.) by using the Extract-n-Amp kit (SigmaAldrich, USA), followed by rolling-circle amplification as previously described (37,38). Amplified DNA was digested with the restriction enzyme BamHI (Fermentas, USA) to generate ϳ2.7-kb monomeric MSV genomes, which were gel purified (GFX; GE Healthcare), ligated into BamHI-digested pGEM-3Zf(ϩ) (Promega, USA) using T4 DNA ligase (Fermentas, USA), and used to transform competent E. cloni cells (Lucigen Corporation, USA).…”
Section: Methodsmentioning
confidence: 99%
“…Viral DNA was isolated from symptomatic leaf material at between 20 and 127 days (median, 47 days) postinoculation (p.i.) by using the Extract-n-Amp kit (SigmaAldrich, USA), followed by rolling-circle amplification as previously described (37,38). Amplified DNA was digested with the restriction enzyme BamHI (Fermentas, USA) to generate ϳ2.7-kb monomeric MSV genomes, which were gel purified (GFX; GE Healthcare), ligated into BamHI-digested pGEM-3Zf(ϩ) (Promega, USA) using T4 DNA ligase (Fermentas, USA), and used to transform competent E. cloni cells (Lucigen Corporation, USA).…”
Section: Methodsmentioning
confidence: 99%
“…Circular viral DNA molecules were amplified from a crude total DNA extract from the two maize samples using Phi29 DNA polymerase (TempliPhi TM , GE Healthcare, USA) as described previously [15,16,18] and cut with KpnI (PR50) and BamHI (PR52) to yield *2.7-kb DNA fragments, which were subsequently ligated to the KpnI and BamHI sites of pGEMZf? (Promega Biotech, USA).…”
mentioning
confidence: 99%
“…To determine the suspected begomoviruses associated with CMD symptoms, leaf samples were collected from five symptomatic cassava plants in Cameroon, and from three symptomatic plants in Togo in August 2014 (Table 1). Total DNA was isolated as described by Shepherd et al (2008). Circular, single-stranded DNA was enriched by rolling circle amplification (RCA).…”
Section: Figurementioning
confidence: 99%