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Stable plasmid clones were obtained from all 244 NFLG-positive PCR products, and both strands of each genome were sequenced, using a primary set of 46 primers.
In all cases both strands of each fragment were sequenced.
Both strands of each fragment were sequenced at least once.
Both strands of each amplicon were sequenced on a 3730 DNA Analyzer (Applied Biosystems).
We sequenced both strands of each product using DNA from all the family members.
Using a set of five PCR fragments covering exons 4 to 10, both strands of each exon were sequenced.
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In addition, when considering both strands of DNA sequences within each group, because l-mers and their reverse complement l-mers (e.g., 4-mers: AAAA/TTTT, GCGC/GCGC, ACCC/GGGT) have the same frequencies, a technique as in [ 20, 28] was used to reduce the size of the vector.
To confirm the identity of the PCR products from the T. vaginalis JT isolate, both strands of two clones for each transposon, chosen at random, were sequenced using the BigDye Terminator mix (Applied Biosystems, Foster City, CA) and run on an ABI 377 sequencer (Applied Biosystems, Foster City, CA).
After cloning 3'RACE products, both strands of DNA were sequenced for each construct.
Both strands of the longest EST representing each one of the 17 validated ORESTES clusters were sequenced.
From each product both strands of eight clones were sequenced, using the ABI prismBigDye Terminator Cycle Sequencing Ready Reaction kit and analysed using 3730 DNA analyser.
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