Genomic DNA (1 μg) from E. Coli strain DH10B was sheared using a Covaris instrument and made into an Illumina–compatible DNA library using the GeneRead Library Prep (I) Kit or a kit from another supplier. The library was eluted in 18 μl Buffer EB and library concentrations were determined by quantitative PCR.
A mixture of highly GC-rich Bordetella pertussis gDNA (GC content 67.7%) and highly AT-rich Streptobacillus moniliformis gDNA (GC content 26.3%) was pooled, made into an Illumina-compatible DNA library using the GeneRead Library (I) Core Kit, and amplified with the GeneRead Library (I) Amp Kit, which contains GeneRead HiFi Polymerase, or a kit from Supplier K. The amplified libraries were sequenced using the Illumina MiSeq instrument, and fidelity and sequence coverage were analyzed using the Galaxy platform. Low error rates and greater cumulated sequence coverage demonstrate that the GeneRead Library (I) Amp Kit provides superior library amplification compared to the kit from Supplier K.
The GeneRead Size Selection Kit effectively removes adapter dimers and adapter monomers following library preparation. A scaled-up image of the above data showing the correct size distribution of Illumina-compatible library fragments following size selection is shown in inset. FU: Fluorescence units.
A mixture of highly GC-rich Bordetella pertussis gDNA (GC content 67.7%) and highly AT-rich Streptobacillus moniliformis gDNA (GC content 26.3%) was pooled, made into an Illumina-compatible DNA library using the GeneRead Library Prep (I) Kit, and amplified with the GeneRead Library Amp (I) Kit, which contains GeneRead HiFi Polymerase, or a kit from Supplier K. The amplified libraries were sequenced using the Illumina MiSeq instrument, and fidelity and sequence coverage was analyzed using the Galaxy platform. The GeneRead Library Prep Kit provided greater sequence coverage in GC- and AT-rich areas of DNA, compared to the kit from Supplier K.
[A] Genomic DNA (50 ng) was sheared using a Covaris instrument and made into an Illumina-compatible DNA library using the GeneRead Library Prep (I) Kit. Sequencing using an Illumina MiSeq instrument revealed a median coverage of 49-fold with uniform coverage distribution. [B] Genomic DNA (1 μg) from E. coli strain DH10B was sheared and used to generate an Ion Torrent-compatible DNA library using the GeneRead Library Prep (L) Kit or a kit from another supplier. After amplification for 10 cycles, both libraries were sequenced on an Ion Torrent PGM instrument and the cumulated normalized coverage was analyzed.
Library Prep (I) Kit uses an optimized one-tube protocol, with fewer cleanup steps and optional high-fidelity library amplification. The hands-on time and total time required for preparation of library DNA is significantly reduced compared to the library preparation system from Supplier I.
The GeneRead Library Prep (L) Kit uses an optimized, one-tube procedure, with fewer cleanup steps and optional high-fidelity library amplification. The hands-on time and total time required for preparation of library DNA is significantly reduced compared to the library preparation system from Supplier L.
