Description |
DLA, a genome walking method, was adapted for sequencing Mu flanking sequences using the 454 FLX platform. Mu flanking sequences from different Mu lines were amplified by a genome walking method, DLA (Liu et al., Genetics, Submitted) and pooled for the 454 pyrosequencing. The primers used to constr .. [more]uct the DNA library are composite primers. DNA barcodes (6 bp) were inserted between the 454 primer A and the Mu-specific primer to allow different input samples that were pooled in the same 454 run to be distinguished after sequencing. The resulting library amplified with Primer 1 and Primer 2 was subjected to 454 sequencing using 454 primer A. Totally, Gl2 and Mu2 samples were subjected to sequencing. Each experiences two technical replication. Each replication was run in the half plate. [less]
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