Run Browser : Browse : Sequence Read Archive : NCBI/NLM/NIH

Run	Spots	Bases	Size	GC content	Published	Access Type
SRR1745785	11.5M	2.2Gbp	1.6G	42.5%	2015-03-13	public

Quality graph (bigger)(smaller)

241

Phred quality score

This run has 2 reads per spot:

̅L=97, σ=12.1, 100%

̅L=96, σ=13.2, 100%

Legend

		Technical read
		Application Read
L=4, 100%		Length is 4, 100% spots contain this read
̅L=165, σ=92.8, 66%		Average length is 165, standard deviation is 92.8, 66% spots contain this read

Experiment	Library Name	Platform	Strategy	Source	Selection	Layout	Action
SRX832022	angustifolius_p27255	Illumina	RNA-Seq	TRANSCRIPTOMIC	unspecified	PAIRED

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RNA isolated with 500 microL Trizol (incubated 15 minutes @ RT) from ~150 mg of matureing seed (20-26 days after anthesis) ground to a fine powder under liquid nitrogen. After centrifugation at 12000g and 4 degrees Celcius for 10 min the supernatant was mixed with 200 microL chloroform and centrifuged again as above. The upper phase was mixed with 300 microL 0.8M sodium citrate/1.2M NaCl and 300 microL isopropanol and incubated on ice for >10 min to precipitate total RNA. RNA precipitate was centrifuged as above and washed twice in 75% ethanol, then dissoved in diethylpyrocabonated-treated water. QC performed via BioAnalyzer and Qubit. 1 microgram of total RNA used to generated TruSeq RNA libraries and samples were pooled on a single lane of Illumina HiSeq1000 generating 100 bp paired-end reads with a ~250 bp insert size. RNA-seq reads were trimmed for low quality <Q30, primer and adaptor sequences via Cutadapt 1.1 (overlap 10, times 3, minimum length 25). Reads paired with discarded trimmed reads < 25 bp were retained as singleton reads.

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Program	Version
Cutadapt	1.1

Biosample	Sample Description	Organism
SAMN03276514 (SRS810336)		Lupinus angustifolius

Bioproject	SRA Study	Title
PRJNA271721	SRP051748	Lupinus spp. seed transcriptome

Abstract:

Comparison of gene expression across multiple isolates of 4 Lupinus spp.

SRA archive data

SRA archive data is normalized by the SRA load process and used by the SRA Toolkit to read and produce formats like FASTQ, SAM, etc. The default toolkit configuration enables it to find and retrieve SRA runs by accession.

Public SRA files are now available from GCP and AWS cloud platforms as well as from NCBI. Access to most data in the cloud requires a user account with the cloud service provider. The user’s account will incur costs for cloud compute or to copy data outside of the specified cloud service region.

Type	Size	Location	Name	Free Egress	Access Type
SRA Normalized	1,590,416 Kb	AWS	https://sra-pub-run-odp.s3.amazonaws.com/sra/SRR1745785/SRR1745785	worldwide	anonymous
SRA Lite	578,598 Kb	NCBI	https://sra-downloadb.be-md.ncbi.nlm.nih.gov/sos5/sra-pub-zq-11/SRR001/745/SRR1745785/SRR1745785.sralite.1	worldwide	anonymous
		AWS	s3://sra-pub-zq-6/SRR1745785/SRR1745785.sralite.1	s3.us-east-1	aws identity
		GCP	gs://sra-pub-zq-102/SRR1745785/SRR1745785.zq.1	gs.us-east1	gcp identity

Original format

The original files submitted to SRA. These files may require specific software to open, read and interpret data.

Type	Size	Location	Name	Free Egress	Access Type
fastq	960,206 Kb	AWS	s3://sra-pub-src-13/SRR1745785/RF3_ACTTGA_L001_R1.fastq.3025cutadapt.fastq.paired.tar.gz	-	Use Cloud Data Delivery
fastq	997,653 Kb	AWS	s3://sra-pub-src-14/SRR1745785/RF3_ACTTGA_L001_R2.fastq.3025cutadapt.fastq.paired.tar.gz	-	Use Cloud Data Delivery

Egress and Access: what does it mean?

Egressterm used by cloud providers to describe cost (charged to the user) of moving data outside of the storage region

Free Egress column indicates where the data can be accessed without an egress charge:

worldwide can be downloaded from anywhere for free
s3.us-east-1 is free to access from machines running in Amazon's us-east-1 region, access from other regions or transport outside of AWS will require paying egress charges
gs.US is free to access from machines running in Google’s gs.US region, access from other regions or transport outside of GCP will require paying egress charges

Access Typedescribes whether a cloud service user account is necessary for data access. "anonymous" access means general public access

Namecolumn provides either a link to free download location at NCBI or a URL for the cloud provider storage location. Either s3:// for Amazon or gs:// for Google storage

Why is SRA data in the cloud?

In order to support large scale (hyper parallel) data analyses SRA data is now available at GCP and AWS with few caveats:

SRA data is copied to the cloud from NCBI. There may be a lag between availability from NCBI and from CSP (cloud service providers)
To access public data user account with the cloud service provider is required. Your account will incur costs for cloud compute and/or to copy data (either archival or results of your comute) outside of the specified cloud service region
Distribution of protected data is signed by NIH account and requires user to operate in the same region as the data

SRA has also begun to provide access to originally submitted source files:

not all files have been validated by SRA
not all files have been copied to cloud locations (recovering it from NCBI tape system takes time ).
the volume of this type of data a much larger and it is not used as often so we will keep most of it on tape or "cold" storage in cloud. As a result the data may not be available instantly and restore requests will be served on first-come first-served basis and cost of resore may be charged to your user account.

What is "Cloud Data Delivery"?: Please visit SRA Data Delivery service to request Sequence Read Archive (SRA) data to be delivered to an Amazon Web Services (AWS) or Google Cloud Platform (GCP) bucket of your choice.

Taxonomy Analysis

Unidentified reads: 94.7%
Identified reads: 5.3%

View in Krona

Strong signals

SuperKingdom	Organism	Rank	%%	Kbp
Eukaryota	Glycine max	species	2.4	54,083
Eukaryota	Arachis	genus	0.7	16,423
Eukaryota	Medicago truncatula	species	0.5	10,682
Eukaryota	Phaseolus vulgaris	species	0.4	9,969
Eukaryota	Cicer arietinum	species	0.4	9,814
Eukaryota	Vigna	genus	0.3	5,942
Eukaryota	malvids		0.3	5,610
Eukaryota	Malpighiales	order	0.1	2,713
Eukaryota	Rosales	order	0.1	2,525

How to read results?: Results show distribution of reads mapping to specific taxonomy nodes as a percentage of total reads within the analyzed run. In cases where a read maps to more than one related taxonomy node, the read is reported as originating from the lowest shared taxonomic node. So when a read maps to two species belonging to the same genus, it is assigned at the genus level. Sequence reads from a single organism will map to several taxonomy nodes spanning the organism’s lineage. The number of reads mapping to higher level nodes will typically be greater than those that map to terminal nodes.

STAT results are proportional to the size of sequenced genomes. Given a mixed sample containing several organisms at equal copy number, proportionally more reads originate from the larger genomes. This means that the percentages reported by STAT will reflect genome size and must be considered against the genomic complexity of the sequenced sample.

How can I query this analysis data?: Please take a look at NCBI SRA Cloud-based Taxonomy Analysis Table

How taxonomy analysis is done?: A fast, scalable, MinHash-based k-mer tool to assess Sequence Read Archive next generation sequence submissions

Lupinus angustifolius cv P27255 Seed RNA-seq(SRR1745785)

Abstract:

SRA archive data

Original format

Taxonomy Analysis

Strong signals

Intensity graph

	View: biological readstechnical readsquality scoresadvanced options Hide Reads base space color space show separated reads quality score with bases show clipped show 4 channel quality scorewith bases Intensity graph Signals table
	Intensity graph