All Questions
15
questions
1
vote
1
answer
40
views
Hybrid assembly versus polishing for hifi and illumina reads
I will have to carry out a project of assembly using hifi reads for which I have already illumina reads and I am wondering which of the hybrid assembly or polishing would be the best option for this ...
- 23
1
vote
1
answer
61
views
How to subset an SRA file for a single chromosome?
I used prefetch to get the Pacbio reads of chicken from the SRA database. I want to align these reads against a reference genome, but not all the reads. I am only interested in a particular region on ...
- 105
0
votes
2
answers
32
views
Assemble Anaeroplasma species genome from metagenomic PacBio data
I have a fasta file containing reads generated by PacBio HiFi whole genome sequencing of a feces sample from mouse.
I would like to use this dataset to generate an assembled circularized genome for an ...
- 1
0
votes
0
answers
222
views
How to manually curate a genome assembly for sequence variation or error?
I have a PacBio HiFi assembly of 1.1 Gb from a heterozygous species. I have aligned this assembly against a reference genome which is around 0.9 Gb. I can see that there are quite a few INDELs, ...
- 75
0
votes
1
answer
74
views
How to improve a genome assembly using Dovetail and PacBio assembly?
I have more of a conceptual question. I have two genome assemblies from the same plant, one from Dovetail technology (~998 Gb) and another is PacBio HiFi assembly (~1.1 Gb). The Dovetail assembly is ...
- 75
0
votes
1
answer
591
views
How to de novo hybrid assemble with Pacbio CCS and Illumina PE reads
I would like to perform de novo genome assembly on a diploid microalgal strain.
I have two datasets:
PacBio CCS/HiFi reads, low coverage.
Illumina PE 2x150 (standard shotgun)
Does anybody have any ...
- 31
0
votes
2
answers
202
views
How do you set the coverage in PacBio's Sequel II?
I am reading the Whole Genome Sequencing for de novo Assembly Best Practices
Use the Sequel II or IIe System and SMRT® Cell 8M to sequence to desired coverage depth for complexity
of genome
10- to 15-...
- 280
1
vote
2
answers
794
views
Is it possible to filter contaminated reads for raw PacBio sequences (not HiFi reads) before assembly?
De novo genome assembly for non-model organisms face the issue of bacterial contamination. For assembled contigs with mostly bacterial-like sequences (based on BLAST search), the entire contig can be ...
1
vote
1
answer
106
views
Gap-fill assembly with PacBio reads
I would like to gap-fill (or correct) a mammalian-sized assembly for which we have BACs that have been sequenced with PacBio CLR data. I have seen there are pbm22/gcpp tools available for correcting ...
- 2,344
2
votes
0
answers
353
views
Platanus-allee phasing fail: Error(13): Error, SolveDBG exception!
I am using Platanus-allee 2.2.2 for heterozygous genome (~500mb) assembly with Illumina short reads and PacBio reads input data.
I have the file contigs.fa from short reads but phasing step with ...
- 21
2
votes
1
answer
70
views
polishing assembled genome to QV50 value
I am trying to assemble some genomes from pacbio reads using HGAP3 followed by quiver polishing. However, even after multiple rounds of quiver polishing, I cannot attain quality value 50 (QV50) for a ...
- 21
2
votes
3
answers
258
views
What assembler is appropriate for High-Fidelity PacBio reads
What assembler is appropriate for High-Fidelity PacBio reads?
For example, canu is good for high-error PacBio reads. But what algorithm to use for HiFi reads? Would it be OK to use canu without the ...
- 2,469
1
vote
1
answer
73
views
How many reads do I need for hybrid assembly
I have Illumina and PacBio reads and I would like to use dbg2olc for hybrid assembly.
Part of dbg2olc is SelectLongestReads which select reads that sum to ...
- 453
8
votes
2
answers
2k
views
estimate genome size: kmer-based approach from PacBio reads
Can anyone suggest a software/method for kmer analysis using PacBio reads (RSII)?
Something similar to Jellyfish, that I saw in a nice tutorial - but must be suitable for long, noisy reads. ...
- 2,706
6
votes
1
answer
76
views
What is the current state-of-the-art in assembling hybrid transcriptomes?
We are considering attempting de novo assembly of a species transcriptomes (i.e. without a reference genome) using the combined NGS outputs of Iso-seq and Illumina.
One example I saw (Li et al 2017),...
- 3,331