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The E. coli DH10B strain was used as the cloning host, and six clones were selected to be sequenced using the Sanger method at MACROGEN (Seoul, South Korea).
With an estimated size of 2.5 Gb, the beetle's genome has been proposed to be sequenced using the novel parallel sequencing technologies including sequencing by synthesis (SBS) such as Illumina®, SOLiD™ and 454 [23].
One of the first nonmodel genomes to be sequenced using the Illumina technology was the giant panda (Li et al. 2010).
As such, we wondered why the PaP1 genome could be successfully sequenced using Roche/454 technique but could not be sequenced using the shotgun method.
This conservativeness makes it possible to design universal primers for amplifying fragments covering almost the whole plastid genome, and the fragments can be sequenced using the Sanger method.
However, despite ETD, some arginine-rich regions, such as the PRM1 core (residues 13 to 41) that contains roughly 75% arginines, could not be sequenced using the bottom-up approach.
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The fragments were sequenced using the automated sequencer 3130 Genetic Analyser (Applied Biosystems, Foster City, CA).
Plasmid DNAs were sequenced using the 3730 DNA Sequencer (Applied Biosystems, Foster City, CA).
Libraries are sequenced using the Illumina MiSeq benchtop sequencer.
The libraries are sequenced using the Illumina HiSeq 2000 Sequencer (BGI, Shenzhen, China).
The PCR products were sequenced using the PRISM Ready Reaction DyeDeoxy Terminator Cycle Sequencing Kit Applied Biosystemss) and an ABI3130 automated sequencer (Applied Biosystems).
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