Sequencing Strain
The parental line of the sequenced aphid clone, LSR1, was
collected in a field of alfalfa (Medicago sativa) near Ithaca, New
York, in 1998 [96]. Aphids for DNA isolation resulted from a
single generation of inbreeding to produce LSR1.AC.G1. The
LSR1.AC.G1 aphid line was grown from a single female and
treated with ampicillin to remove R. insecticola. Prior to DNA
preparation, aphids were heat treated to reduce the number of
Buchneracells; entire aphid colonies on broad bean plants were
placed in a 30uC incubator for 4 d. RT qPCR quantification of
Buchnera/aphid DNA ratios revealed a significant decrease in the
level of Buchnera relative to aphids not subjected to heat.
Approximately 2% of the sequencing reads came from the
Buchneragenome and were removed for separate assembly of
Buchneragenome.