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In ten of the areas PCR and sequencing was carried out to validate the sequence.
This integrated alignment was used to assess and validate the sequence assemblies and to build the physical map further.
In order to validate the sequence of the wild-type strain and to determine the complete MDR and XDR sequences, we independently sequenced three KZN strains (one wild type, one MDR, and one XDR) using whole-genome sequencing.
The identified positions were then manually validated on the electropherogram to validate the sequence.
Forward and reverse sequencing was also performed to validate the sequence.
To validate the sequence assembled by Phrap, the 7096 traces were reassembled with the Celera WGS assembler [ 43].
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Finally, we applied a series of functional assays to validate the sequencing results.
To validate the sequence-specificity of gene knock-down in vitro, after transfection the fibroblasts were analyzed for protein and mRNA downregulation by immunoblots and real-time RT-PCR.
These results fully validate the sequencing eletropherogram measurement as an accurate method to determine mA%.
The efficient and powerful approach developed herein can be successfully used to validate the sequences of miRNAs, especially the termini, which depict the complete miRNA sequence in the computationally predicted precursor.
All vectors were sequenced to validate the sequences.
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