Tools for Reference-Guided Assembly Using Nanopore Sequencing Reads
4
0
Entering edit mode
4 weeks ago

I have raw reads generated from Nanopore sequencing and have already performed a de novo assembly. Now, I would like to perform a reference-guided assembly using these reads. My questions are:

What are the recommended tools for performing reference-guided assembly with Nanopore reads? Are there any specific considerations or best practices for using these tools with Nanopore sequencing data?

Assembly Nanopore Reference-guided Genome-assembly Tools • 506 views
ADD COMMENT
1
Entering edit mode
4 weeks ago
Michael 55k

I think you should look into RagTag. Considerations regarding the use of reference-based correction and scaffolding depend on the field of application, the similarity and quality of the reference genome, and which ragtag steps you would like and can afford to apply. I would advise caution for over-correction. If you apply all steps, the outcome will be 99.9% identical to the reference, then, why sequence in the first place? If you want to look into structural variants or SNVs, or have haplotype-resolved assemblies, I would not apply reference-based corrections.

ADD COMMENT
0
Entering edit mode
4 weeks ago

I would not do reference guided assembly with nanopore ( to be clear assembling reads into contigs using a related reference) and I do not know of any tools which exist.

As Michael says, instead use RagTag to map your assembled contigs to a closely related reference if you want to assemble to chromosome level.

ADD COMMENT
0
Entering edit mode

ok i understand. Thank you for your suggestion.

ADD REPLY
0
Entering edit mode
4 weeks ago

if you consider to ignore the advice of Michael and colindaven (and their arguments why not to) , you can have a look at flye or rebaler , I think they can use a reference in the assembly process.

ADD COMMENT
0
Entering edit mode
4 weeks ago
Umer ▴ 130

SPAdes also has a --known-contigs option where you can input a reference genome.

Spades has an option for Nanopore data but I haven't seen any Nanopore assembly by spades.

ADD COMMENT
1
Entering edit mode

You should probably try RagTag for your own genomes that you have posted in other threads.

ADD REPLY

Login before adding your answer.

Traffic: 2562 users visited in the last hour
Help About
FAQ
Access RSS
API
Stats

Use of this site constitutes acceptance of our User Agreement and Privacy Policy.

Powered by the version 2.3.6