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Library amplification readymix

Manufactured by Roche

The Library Amplification Readymix is a pre-formulated reagent solution designed for amplifying DNA libraries. It contains the necessary enzymes, buffers, and dNTPs required for the library amplification step in next-generation sequencing workflows.

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3 protocols using library amplification readymix

1

Multiplexed Paired-end Library Construction

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Multiplexed paired-end libraries were constructed from 5 μg of genomic DNA purified using the Purgene kit (Qiagen). The genomic DNA was sheared by sonication and end-repaired using the End-it DNA End-repair kit (Epicentre Technologies). Common adaptors from the Multiplexing Sample Preparation Oligo Kit (Illumina) were then ligated to the genomic DNA fragments, and the fragments were then subjected to 18 cycles of amplification using the Library Amplification Readymix (KAPA Biosystems). The amplified products were fractionated on an agarose gel to select 600 bp fragments, which were subsequently sequenced on an Illumina HiSeq 2000 using the Illumina GAII sequencing procedure for paired-end short read sequencing. Reads from each read pair were mapped separately by bowtie version 2.2.1 [108 (link)] to a reference sequence that contained revision 64 of the S. cerevisiae S288c genome [109 (link)], hisG from Samonella enterica, and the kanMX4 marker (S3 Table). Reads are available from National Center for Biotechnology Information Sequence Read Archive under accession number: SRP107803.
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2

Multiplexed Yeast Genome Sequencing

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Multiplexed paired-end libraries were constructed from 5 µg of genomic DNA purified using the Purgene kit (Qiagen). The genomic DNA was sheared by sonication and end-repaired using the End-it DNA End-repair kit (Epicentre Technologies). Common adaptors from the Multiplexing Sample Preparation Oligo Kit (Illumina) were then ligated to the genomic DNA fragments, and the fragments were then subjected to 18 cycles of amplification using the Library Amplification Readymix (KAPA Biosystems). The amplified products were fractionated on an agarose gel to select 600 bp fragments, which were subsequently sequenced on an Illumina HiSeq 2000 using the Illumina GAII sequencing procedure for paired-end short read sequencing. Reads from each read pair were mapped separately by bowtie version 0.12.7 [74] (link) to a reference sequence that contained revision 64 of the S. cerevisiae S288c genome (http://www.yeastgenome.org), hisG from Samonella enterica, and the hphMX4 marker (Table S1). Sequencing data have been deposited at NCBI Sequence Read Archive (http://www.ncbi.nlm.nih.gov/sra) under the accession SRP039033.
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3

Multiplexed Paired-end Library Construction

Check if the same lab product or an alternative is used in the 5 most similar protocols
Multiplexed paired-end libraries were constructed from 2 μg of genomic DNA purified using the Purgene kit (Qiagen). The genomic DNA was sheared using M220 focused-ultrasonicator (Covaris) and end-repaired using the End-it DNA End-repair kit (Epicentre Technologies). Common adaptors from the Multiplexing Sample Preparation Oligo Kit (Illumina) were then ligated to the genomic DNA fragments, and the fragments were then subjected to 18 cycles of amplification using the Library Amplification Readymix (KAPA Biosystems). The amplified products were fractionated on an agarose gel to select 600 bp fragments, which were subsequently sequenced on an Illumina HiSeq 4000 using the Illumina GAII sequencing procedure for paired-end short read sequencing. Reads from each read pair were mapped separately by bowtie version 2.2.1 [72 (link)] to a reference sequence that contained revision 64 of the S. cerevisiae S288c genome (http://www.yeastgenome.org), hisG from Samonella enterica, and the hphMX4 marker. Sequence data is available from National Center for Biotechnology Information Sequence Read Archive under accession number: SRP106876.
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