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_14
Snippet: To analyze the lineages of TEV-ÃNIb and TEV evolved in N. tabacum 35S::NIb (second experiment above), total RNA was extracted using RNAeasy Plant Mini Kit (Qiagen) from symptomatic leaves of five N. tabacum 35S::NIb infected by the evolved lineages of TEV and five N. tabacum 35S::NIb infected by the evolved lineages of TEV-ÃNIb. To verify the presence of deletions, the full genome of each genotype was reverse transcribed using M-MLV RT (Ferment.....
Document: To analyze the lineages of TEV-ÃNIb and TEV evolved in N. tabacum 35S::NIb (second experiment above), total RNA was extracted using RNAeasy Plant Mini Kit (Qiagen) from symptomatic leaves of five N. tabacum 35S::NIb infected by the evolved lineages of TEV and five N. tabacum 35S::NIb infected by the evolved lineages of TEV-ÃNIb. To verify the presence of deletions, the full genome of each genotype was reverse transcribed using M-MLV RT (Fermentas) and primer 97-101-R (supplementary table S1, Supplementary Material online), then PCR amplified using the Ultra High-Fidelity DNA polymerase Phusion (Finnzymes) and primers 73-80-F and 97-101-R (supplementary table S1, Supplementary Material online). Deletions were confirmed by resolution of PCR products on 1% agarose gels.
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