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how to add inline barcode to raw DNA fragment without PCR

Dear all,

I want to use inline barcodes (barcode adjacent to the DNA fragment, included as the starting of the sequencing reads) for my illumina sequencing. Then I found most of current posts used this technique for short amplicon sequencing (e.g. 16S sequencing), in which the inline barcodes can be included during the PCR amplification. However, in my situation, I can only add the inline barcode after PCR and fragmentation, so I wonder if there is any kit or suggestion to add the inline barcode to the end-prep DNA fragment (I think shoule be done by ligation), or where I can design and synthesize the inline barcodes (essentially, short double strand DNA)?

Thanks.

next-gen sequencing illumina
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updated by Biostar Biostar • written by liaoyunshi • liaoyunshi

This forum is mainly for bioinformatics discussion while your question seems to be about barcodes. Are you interested in software to design barcodes, or about laboratory strategies or kits?

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written by Sean Davis Sean Davis

You should search SeqAnswers.com for similar questions. This question may also be more appropriate for that forum.

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written by GenoMax GenoMax

As you mentionned it in your question this type of issue can be solved by using a ligase (i.e. T4 ligase) with or without a preliminary restriction enzyme step. Design can be done by hand since the oligo is relatively simple. Synthesis depends on the length of your fragment, most compagnies selling custom oligonucleotides offer a broad range of size. Only trick is to make sure you design it in the right orientation (sequencing requires a 5'==>3' orientation).

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written by VHahaut VHahaut

Hello liaoyunshi!

We believe that this post does not fit the main topic of this site.

Not related to bioinformatics

For this reason we have closed your question. This allows us to keep the site focused on the topics that the community can help with.

If you disagree please tell us why in a reply below, we'll be happy to talk about it.

Cheers!

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written by WouterDeCoster WouterDeCoster

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