Question: (Closed) how to add inline barcode to raw DNA fragment without PCR
gravatar for liaoyunshi
14 months ago by
liaoyunshi0 wrote:

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)?


sequencing next-gen illumina • 506 views
ADD COMMENTlink modified 13 months ago by Biostar ♦♦ 20 • written 14 months ago by liaoyunshi0

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?

ADD REPLYlink written 14 months ago by Sean Davis25k

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

ADD REPLYlink written 14 months ago by genomax63k

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).

ADD REPLYlink modified 14 months ago • written 14 months ago by VHahaut1.1k

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.


ADD REPLYlink written 13 months ago by WouterDeCoster37k
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