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  • vl80
    Member
    • Nov 2012
    • 34

    #1

    sequencing 1000 amplicons on miseq

    Hi,

    We are developing an amplicom sequencing project where the consensus primers for enrichment can be designed with amplicons between 800 to 1100 bp only. However, there is another catch, we do not want to use a single Illumina index per sample as this would increase the expenses. Previously we successful developed a similar method where in house nucleotides were added to the primers and several samples were pooled together to create and "Index" sample that would be library prepped with a Truseq kit.
    However, as the amplicons are longer this time, and as we need the whole length of the amplicon to be sequenced, we will have to fragment the amplicons. Is there a method to add 5 nt barcodes to the fragmented products that is primer independent?
    Thanks a lot

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