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All PCR products were sequenced and samples with overlapping sequencing traces were subcloned and individual alleles determined.
The cpn60 UT is accessible in nearly all bacterial genomes with a single set of universal primers, and its length is such that it can be completely sequenced in one pair of overlapping sequencing reads via di-deoxy sequencing.
There is some hope that complete mtDNA sequencing could reach a much higher quality level (by perhaps two orders of magnitude) when stringent lab routines are followed and overlapping sequencing of both strands is applied, such as may have been achieved with the data produced by [8].
Both strands were sequenced by primer walking with overlapping sequencing reactions.
Electropherograms of overlapping sequencing fragments were manually edited using the software package Sequence Navigator version 1.1 from PE Biosystems.
Electropherograms of overlapping sequencing fragments were manually edited using the software Sequence Navigator version 1.1, from PE Biosystems.
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Overlapping sequences that were not identical in the overlapping regions were not removed since they might contain different domains.
Each primer pair contains non-overlapping sequences at their 3' end and primer-primer complementary (overlapping) sequences at the 5' end.
After the sequence of each fragment is determined, powerful computers assemble the pieces in the correct order by looking at overlapping sequences.
The point of view shifts constantly, and overlapping sequences fade in and out like dreams.
The semi-quantitative technique is based on the amplification of short, overlapping sequences differing in size.
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