Long read sequencing reveals poxvirus evolution through rapid homogenization of gene arrays
Dr. Brian Bodemann, Account Manager, West «Advancing QC for
long read sequencing with the FEMTO Pulse»
The PromethION will produce long reads on an industrial scale, meaning it will be a platform in high demand for genome assembly and transcriptomics work as well as population - scale
long read sequencing.
Here, we describe the results obtained from our current
long read sequencing project of 100 Icelanders with Oxford Nanopore technology.
Not exact matches
Until now, such large - scale data storage has been impractical because of the difficulty and cost of
reading and writing
long sequences of DNA.
[Volodymyr Kuleshov et al, Synthetic
long -
read sequencing reveals intraspecies diversity in the human microbiome]
But short
reads can be hard to interpret during the overlaying process and there hasn't been a way to
sequence long fragments of DNA in a targeted and more efficient way.
«By reducing non-Y chromosome
reads from our data with flow sorting and the RecoverY technique that we developed, and by using this combination of
sequencing technologies, we were able to assemble the gorilla Y chromosome so that more than half of the
sequence data was in chunks
longer than about 100,000 bases in length,» said Medvedev.
Many of these repeats, including some genes, appear as back - to - back series of the same repeated
sequence or as
long palindromes which, like the word «racecar,»
read the same forward and backward.
To their surprise and dismay, the genome turned out to contain
long stretches of noncoding and repetitive DNA, which made it difficult to piece together the short
reads produced by the
sequencing machine.
The key to their success was to generate
long sequencing reads, producing an assembly with not many more scaffolds than C. krusei has chromosomes.
To overcome the extreme genomic complexity, the team used new
long -
read sequencing technology that boosted the quality of the genome
sequence obtained by more than one hundred fold over standard short -
read approaches.
DNA
sequencing and genome mapping can thus be compared to dividing a very
long text into lots of small pieces that are
read separately — letter by letter, or more exactly: nucleobase per nucleobase.
A new
sequencing technology based on
longer sequence reads allows missing genes and missing forms of genetic variation to be discovered for the first time.
The researchers used Single Molecule, Real - Time (SMRT)
sequencing technology, the assembly tools Falcon and QUIVER, and other techniques to generate
long sequence reads.
Current technology for «
long read» detailed genomic
sequencing can be performed using expensive instrumentation (around # 500,000).
Eichler predicted, «In five years there might be a
long -
read sequence technology that will allow clinical laboratories to
sequence a patient's chromosomes from tip to tip and say, «Yes, you have about three to four million SNPs and insertions deletions but you also have approximately 30,000 - 40,000 structural variants.
The SMRT technology used in the new study makes it possible to
sequence and
read DNA segments
longer than 5,000 bases, far
longer than standard gene
sequencing technology.
This study aimed to assess the feasibility of using the Oxford Nanopore Technologies (ONT) MinION
long -
read sequencer in reconstructing fully closed plasmid
sequences from eight Enterobacteriaceae i...
Recent and rapid advances in technologies that permit large - scale creation and synthesis («writing») of
longer pieces of synthetic DNA, as well as the advent of extremely fast, cheap and accurate
sequencing («
reading») of DNA, have changed our collective thinking about the feasible size and scope of projects in many labs.
We describe the concordance between the different
sequencing platforms both for variant and methylation calling and describe some of the additional information provided by the
long read data.
An open source computer program for ordering short DNA
sequence reads into
long DNA fragments with the aid of the barcodes is also provided.
The recently launched Gemcode system adds a new dimension to short -
read DNA
sequencing, by enabling the ordering of millions of barcoded short
sequence reads of 150 base pairs into
long continuous DNA molecules of hundreds of thousands of base pairs in size.
To meet the challenge they used «next - generation»
sequencing techniques, in which the DNA is broken up randomly into numerous small segments and assembled into
longer sequence reads by identifying the overlapping ends.
Short DNA fragments are not able to reassemble repetitive elements in the genome — for these hard to assemble regions
longer sequence read information is needed.
Stephen R. Quake, Ph.D., Stanford University, Palo Alto, Calif. $ 1.8 million (3 years) «High - Throughput, Single - Molecule DNA
Sequencing» This group will try to improve its sequencing - by - synthesis technology in order to achieve longer reads from very large numbers of single DNA
Sequencing» This group will try to improve its
sequencing - by - synthesis technology in order to achieve longer reads from very large numbers of single DNA
sequencing - by - synthesis technology in order to achieve
longer reads from very large numbers of single DNA molecules.
DNA sequencers can not
sequence long pieces of DNA, so the extracted DNA molecules were sheared to different sized fragments in a controlled manner such that when they are
sequenced they can be reassembled computationally by matching overlapping codes in the
reads — creating
long sequences of the genetic code.
- Scientists have
long been
reading the code of life — the genome — , as a
sequence of letters but now researchers have also started exploring its three - dimensional organisation.
The method enables use of widely available short -
read DNA
sequencing platforms to study
long single molecules within a complex sample, without losing information of physical connection between
sequencing reads from each molecule of origin.
New generation of
sequencing technology uses nanopores to deliver ultra
long read length single molecule
sequence data, at competitive accuracy, on scalable electronic GridION platform.
With
longer (> 50 bp) fragment - end
reads and / or paired - end libraries, it's possible to detect small insertion / deletion variants (indels) in next - gen
sequencing data.
Fortunately, indel detection is one area that will be helped dramatically by improvements to the
sequencing technologies, namely
longer reads and paired - end protocols.
Corresponding advances in
long -
read SV calling algorithms have reduced coverage requirements, making
long -
read genome
sequencing a cost - effective approach for both disease research and population genetics studies.
Alexander Hoischen, Ph.D., Radboud University Medical Center «Medical genetics: Identification of hidden structural variants with
long -
read sequencing»
The
long sequencing reads produced by Oxford Nanopore's platforms enable the assembly of genomes with superior contiguity compared to those produced by second generation technologies.
In these cases,
longer reads are necessary to reconstruct the entire
sequence.
Nanopore
sequencing is a promising technique for genome
sequencing due to its portability, ability to
sequence long reads from single molecules, and to simultaneously assay DNA methylation.
Thursday, Oct. 19, 4:15 - 6:15 p.m., Room 310A, South Building Invited Session: Analysis of Cancer Genome Variation Using
Long -
read Sequencing Moderators: Jeffrey Rosenfeld, Rutgers Cancer Institute; and Sara Goodwin, Cold Spring Harbor Laboratory
Current research projects include
sequencing the wheat genome through a combination of Illumina short
read sequencing and
long sequence reads using the new Pacific Biosciences sequencers.
Speed, single - base sensitivity and
long read lengths make nanopores a promising technology for high - throughput
sequencing.
Compared with short
reads, the assemblies obtained from
long -
read sequencing platforms have much higher contig continuity and genome completeness as
long fragments are able to extend paths into problematic or repetitive regions.
Long -
read sequencing technologies are transforming our ability to assemble highly complex genomes.
Here, we employed cDNA amplicon
sequencing using a
long -
read portable sequencer, MinION, to characterize various types of mutations in cancer - related genes, namely, EGFR, KRAS, NRAS and NF1.
The MinION is a portable single - molecule DNA
sequencing instrument that was released by Oxford Nanopore Technologies in 2014, producing
long sequencing reads by measuring changes in ionic flow when single - stranded DNA molecules translocate through the pores.
Unlike other methods, which
read chunks of DNA perhaps a hundred or so base pairs in length, the MinION can
read sequences as
long as 882,000 base pairs in length using the nanopore technology.
To accelerate lncRNA annotation, the GENCODE consortium has developed RNA Capture
Long Seq (CLS), which combines targeted RNA capture with third - generation long - read sequenc
Long Seq (CLS), which combines targeted RNA capture with third - generation
long - read sequenc
long -
read sequencing.
Long -
read sequencing technologies were launched a few years ago, and in contrast with short -
read sequencing technologies, they offered a promise of solving assembly problems for large and complex genomes.
27/2: 30
Long read single - molecule real - time (SMRT) full gene
sequencing of cytochrome P450 2D6 (CYP2D6).
Long - read sequencing technologies such as Pacific Biosciences and Oxford Nanopore MinION are capable of producing long sequencing reads with average fragment lengths of over 10,000 base - pairs and maximum lengths reaching 100,000 base - pa
Long -
read sequencing technologies such as Pacific Biosciences and Oxford Nanopore MinION are capable of producing
long sequencing reads with average fragment lengths of over 10,000 base - pairs and maximum lengths reaching 100,000 base - pa
long sequencing reads with average fragment lengths of over 10,000 base - pairs and maximum lengths reaching 100,000 base - pairs.
Rapid low - cost assembly of the Drosophila melanogaster reference genome using low - coverage,
long -
read sequencing