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Another potential source of error in distance estimations arises during sequencing.
The reverse primer included a 5 bp barcode for multiplexing of samples during sequencing.
Mixtures at both of these levels are distinguishable from non-heteroplasmic during sequencing.
Custom barcoded adaptor oligos (IDT) were used in step 3.9.2 to enable pooling multiple libaries in a single emulsion PCR reaction and picotiter plate region during sequencing.
Custom barcoded adaptor oligos (IDT) were used to enable pooling multiple libaries in a single emulsion PCR reaction and picotiter plate region during sequencing.
The sequences shown do not include adaptor or barcode sequences, and the reverse primer included a 5 bp barcode for multiplexing of samples during sequencing.
Thus these revised criteria have considerably improved our specificity and reduced artifacts likely arising from errors in reverse transcription that are subsequently over-sampled during sequencing.
Default parameters were used at all instances during sequencing.
Real-time monitoring was performed for each cycle during sequencing.
The gaps had G+C rich regions that resulted in early signal loss during sequencing.
This indicates that very specific base pairing is highly prone to error during sequencing.
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