Navigation Links
No need to prepare
Date:12/11/2012

For the first time, researchers sequenced DNA molecules without the need for the standard pre-sequencing workflow known as library preparation.

Using this approach, the researchers generated sequence data using considerably less DNA than is required using standard methods, even down to less than one nanogram of DNA; 500 times less DNA than is needed by standard practices.

Libraries are collections of DNA fragments derived from genomic samples using molecular biology techniques specific to the sequencing technology being used. They are laborious, time consuming, and often DNA consuming. This new technique has the potential to greatly reduce DNA consumption and the time it takes to generate sequencing data from small genomes.

"This is the first time that anyone has been able to directly sequence single molecules of DNA in this way." says Dr Paul Coupland, first author from the Wellcome Trust Sanger institute. "We applied our approach to sequencing the genomes of viruses and bacteria and found that even with a relatively low level of optimisation, we were able to determine what organisms we were looking at, whether specific genes or plasmids were present in the sample (which can be important in determining antibiotic resistance) as well as other information such as modifications to specific DNA bases.

"Once optimised, our technique has great potential for a fast and efficient way of identifying organisms in hospitals and other healthcare settings. It also gives us the absolute confidence there is no library based bias in the sequence data, quite simply because no library was ever created."

The team utilised the PacBio RS, a sequencing system that sequences single molecules of DNA, for the direct sequencing approach. Sequence data was generated from small circular single-stranded and double-stranded DNA viral genomes, as well as from linear fragments covering the entire genome of an MRSA strain of Staphylococcus aureus.

The team tried analysing the genome of one organism using only eight hundred picograms of DNA, over six hundred times less than the quantity used in standard practice. In this example, the PacBio only generated 70 reads, or fragments of sequence, from the genome. Although this is a fraction of the number of reads generated relative to standard library methods, it was still enough information for the team to identify the specific organism being sequenced; this work could allow the identification of organisms in metagenomic samples that were previously undetectable.

"To sequence microorganisms, one needs to be able to grow them in a lab first," says Dr Tamir Chandra, lead author from the Babraham Institute. "Not only is this time consuming, but sometimes micro-organisms do not grow, making it extremely difficult to sequence their genome.

"With this method we can directly sequence these organisms and find out their identity in a short space of time."

"Our role at the Sanger Institute is to determine how we can utilise and improve these sequencing platforms to generate biological information more efficiently and in turn, possibly, influence the control and treatment of disease and infections." says Dr Harold Swerdlow, lead author from the Wellcome Trust Sanger Institute. "Our technique can be performed without any prior knowledge of the sequence and with no organism specific reagents, in a short space of time. This makes it a promising alternative for clinical situations such as infection control."


'/>"/>

Contact: Aileen Sheehy
press.office@sanger.ac.uk
44-012-234-92368
Wellcome Trust Sanger Institute
Source:Eurekalert

Related biology news :

1. Lizard moms may prepare their babies for a stressful world
Post Your Comments:
*Name:
*Comment:
*Email:
(Date:5/16/2017)... , May 16, 2017  Veratad Technologies, LLC ( ... online age and identity verification solutions, announced today they ... Conference 2017, May 15 thru May 17, 2017, in ... and International Trade Center. Identity impacts ... and in today,s quickly evolving digital world, defining identity ...
(Date:5/6/2017)... SINGAPORE , May 5, 2017 ... has just announced a new breakthrough in biometric ... that exploits quantum mechanical properties to perform ... new smart semiconductor material created by Ram Group ... across finance, entertainment, transportation, supply chains and security. ...
(Date:4/19/2017)... 2017 The global military biometrics ... marked by the presence of several large global players. ... five major players - 3M Cogent, NEC Corporation, M2SYS ... nearly 61% of the global military biometric market in ... global military biometrics market boast global presence, which has ...
Breaking Biology News(10 mins):
(Date:8/15/2017)... , ... August 15, 2017 , ... The Conference ... on Immuno-Oncology 360° (IO360°) programming through a series of upcoming panels and events. The ... 7-9, 2018, at The Roosevelt Hotel in New York City. , “With our experience ...
(Date:8/14/2017)... York, NY (PRWEB) , ... August 14, 2017 , ... ... in Clinical Trials event, which will take place on September 6, 2017 at ... Daniel Karlin, MD , Head of Experimental Medicine, Informatics, and Regulatory Strategy, Pfizer Innovative ...
(Date:8/10/2017)... ... 09, 2017 , ... As a full-service marketing agency, 212 ... with the right message. Their effective, cutting-edge inbound marketing strategies are available to ... crucial the agriculture industry is,” said David Phelps, chief marketing officer at 212 ...
(Date:8/10/2017)... ... August 09, 2017 , ... SPIE, the international society for ... Wellman Center for Photomedicine, the Manstein Lab in the Cutaneous Biology Research Center ... at University of California, Irvine — and the Hillenkamp family to establish the ...
Breaking Biology Technology: