Abstract
Here we present the devised BC-store–a program for analyzing and selecting sets of barcodes for sequencing on platforms manufactured by MGI Tech (China). The app is available as an open source in Python3 and as a desktop version. The application allows analyzing the compatibility of barcodes on a single lane of a flow cell in a set in the case of equal and arbitrary fractions. In addition, with the help of this tool barcodes can be added to an existing set with custom share options. In this paper we describe how BC-store works for different tasks and consider the effectiveness of using BC-store in sequence lab routine tasks.
Highlights
To pool multiple samples on a single lane of the flow cell, barcoding is used–adapters carrying a unique nucleotide sequence are introduced by ligation [1]
The task of selecting the adapters for a set is similar to sequencing low diversity libraries [4]: if all adapters on the lane have the same nucleotide during this sequencing cycle, the quality of its reading drops dramatically
The sets offered by MGI Tech [5] are far from routine practice, as they do not allow for nonequimolar sample pooling by default, and they have other disadvantages
Summary
The barcode for MGISEQ-2000/DNBseq-G400 is a sequence of 10 nucleotides. Several barcodes can correspond to a single sample, but each barcode is associated with only one sample. MGI Tech offers 96 variants of barcodes and a scheme for their optimal combination (Fig 1). This scheme of set combinations has a number of limitations: it is not possible to merge barcodes in different proportions; a certain number of samples are supposed to be used, which limits the opportunities of research. It can lead to uneven and inefficient use of barcodes. According to our subjective perception, MGISEQ-2000 is more sensitive to the correct balance of barcodes in a set than the Illumina HiSeq 2500 that we use as well
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