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Because tandemly duplicated genes tended to be accidentally combined into one gene during alignments, we divided these regions such that each region contained only one candidate gene and aligned the FLcDNAs to each region (Supplementary Fig. S1).
To examine whether both alleles at heterozygous sites were equally considered during alignments, we also created a "complement" reference sequence by swapping the allele at each heterozygous site in the initial custom reference with the alternate allele from the input file.
Similar(58)
Allowing only a single mismatch during alignment prevents reads from aligning if both alleles are not considered.
In cases where the error rate of the sequencing platform exceeds the number of mismatches allowed during alignment, false positive uniquely aligning tags will occur (Supplementary Figure S1).
Because it is preferable to take the amino acid level into account during alignment of protein-coding DNA, the aligned amino acid sequences were used as a scaffold for constructing the corresponding nucleotide sequence alignment using RevTrans 1.4 [ 68].
As common aligners can allow a small amount of mismatches during alignment, reads containing SNPs may still be mapped to the reference.
Both these factors contributed to difficulties during alignment, because a dual class such as 'thoracic segment' in HAO could be aligned with either 'thoracic segment' in FBbt (the general class) or 'adult thoracic segment' in FBbt (the stage-specific class).
To avoid frameshift errors in the nucleotide sequences during alignment step, the deduced amino acid sequences of the protein-coding genes were aligned and subsequently retranslated using ClustalW implemented in BioEdit v.7.1.11 (Hall 1999).
B, C Since the comparison metric most commonly used in SPT is cross correlation, greater density overlap tends to increase the similarity score during alignment.
(In order to maintain time-framing during alignment, data points are missing in some IFs) (Supplementary Figure 2.doc, 56 K. http://www.ejnmmires.com/imedia/1153689202550379/supp2.doc).
In addition, very accurate preparations of parts are required as no gap or in some cases only a minimal gap is permissible during alignment of the parts.
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