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Reference sequence to be used for assembly in nanopore

Hello,

I am a newbie in NGS data analysis. I wanted to analyse the Oxford nanopore's reads. For assembly I am using minimap tool. A reference file is needed for the process. Can I use any related/similar fasta sequence as a reference file or I must use only the Refseq DNA file? Need some help!

reference assembly

Hi Sudosundu,

please include which species you are working on. Is it human? Just towards terminology, minimap is an aligner so it matches DNA sequences to a reference. Assembly would be to put the raw reads together (=assemble) a new reference/contig/genome/whatever-you-sequenced. What do you mean by Refseq DNA file, can you give a link?

Hi Sorry for the less information. I am working on plant virus. My aim is to characterize the viral genome. Refseq DNA file means reference genome to be used for mapping the reads. https://www.ncbi.nlm.nih.gov/refseq/

I wanted to analyse the Oxford nanopore's reads.

It is usually helpful to mention what your final aim is, for us to put you on the right track.

Hi, I just came through your Nanopack. Thanks for the pack. My final aim is to characterise the plant viral genome. I am sequenced virus infected plant samples and to characterize them based on a reference viral genome.

Then you need an assembler, not an aligner.

Do a bit of googling around canu and it's brethren to get you started. You won't need a reference sequence for this.

Can I use any related/similar fasta sequence as a reference file or I must use only the Refseq DNA file?

You could use a related reference but you will need to keep in mind that depending on how different that strain/species is from your organism the results may be less than optimal. That said, if a RefSeq genome is available then that is your best bet for use as a reference for alignment (though it could also be used for a reference guided assembly).

Now I am using the reference genome from the NCBI.

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