Thule, Greenland – Day Three

Day three started with me missing breakfast. It seems that folks around here only eat breakfast between 5am and 8am. Today was a very rough day for sampling.  About an hour drive to the area near the site, about a three-mile hike to one spot another half-mile hike to another spot followed by the three and a half mile hike back to the truck. We sampled “rich” soil and “rich” soil from a lake. These two sites were sampled and categorized as “rich” due to the abundance of vegetation around and near the sites. The area surrounding Thule is very desolate so I can imagine the plants have a hard enough time growing.  It would be very interesting to see what microbes are present in these two sites to allow such vegetation to grow; even more interesting to see how water affects the microbial population. Samples were frozen once we got back to the on site lab. A small portion was saturated with AllProtect to ensure preservation of RNA for transcriptomics analysis.

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The day ended with a lecture from another NSF grant recipient to install a telescope on the Greenlandic ice cap. It was an interesting idea to coordinate radio imaging from other telescopes around the world to look at quantum singularities that were very far away. After speaking to some of the other scientists here I found out that our group, which includes myself and our collaborators Slava Epstein and Dawoon Jung, were the ONLY Microbiologists on the base. Everyone else was either a Geologist, Environmental Scientist, Astronomer, or Meteorologist. It was great to hear about everyone else’s projects.

The Next Generation Science Standards are Ready for Review

The second draft is ready for public comment through January 29th. Please be sure to take some time to review.

http://www.nextgenscience.org/next-generation-science-standards

Plant Bioinformatics Workshop

JCVI recently held its 3rd Annual Plant Bioinformatics Workshop from July 15-19th. During the week-long workshop, 20 scientists from the Plant Research community visited JCVI and learned many aspects of Bioinformatics from the members of Chris Town’s Plant Genome group. Attendees included undergraduate and graduate students, post-doctoral fellows, research scientists and faculty at various Universities throughout the United States as well as a biotech company. In addition to the on-site participants, we had 5 additional participants attend the workshop via WebEx. The virtual participants had the opportunity to sit in on the lectures and complete the hands on exercises by logging into an Amazon Cloud instance, which was set up specifically for this purpose. The topics covered during the workshop included UNIX tools for Bioinformatics, Genome Assembly, Structural and Functional Annotation, RNA-seq assembly and analysis and SNPs. In addition to JCVI’s instructors, we had additional sections covered by external instructors. Eric Lyons (University of Arizona and iPlant) presented on Comparative Genomics and the iPlant Infrastructure and Ann Loraine (UNC Charlotte) presented on Integrated Genome Browser. All sessions contained a hands-on component so the students would have the opportunity to use the tools that we discussed during the lecture portion.  Watch our website for future offerings!

JCVI-1

Carl Woese 1928-2012

Editor’s Note: This post originally appeared on T. Taxus, December 31, 2012, by Jonathan Badger. Dr. Badger  is an Assistant Professor in the Microbial and Environmental Genomics Group at the J. Craig Venter Institute in La Jolla, CA. Reprinted by permission.

As you may have heard, Carl Woese died of pancreatic cancer yesterday at the age of 84. I had the honor of working with Carl in grad school at the University of Illinois where my advisor, Gary Olsen, ran a joint lab with Carl.

Carl Woese

Carl Woese. Photo courtesy IGB.

As the originator of the use of ribosomal RNA to distinguish and classify organisms (including obviously the Archaea), Carl both revolutionized evolutionary biology and created a method that is still very much in use today. Even in the latest metagenomic study of the oceans or of the human gut, a 16S rRNA diversity study is required as a control in addition to whatever additional markers or random sequencing is used.

One of the things that fascinated me about Carl is how he constantly reinvented himself and explored new fields of biology — his early work in the 1960s dealt with classical molecular biology and the genetic code (the origins of which continued to fascinate him for the rest of his life). He then transfered to the study of the ribosome and its structure, which in turn led to his study of 16S and its evolutionary implications. In the 1990s, when I worked with him, he was a pioneeering microbial genomicist and collaborated with TIGR to sequence the first two Archaeal genomes. And in his final years he focused on early evolution and the last common ancestor of life in the light of what genomics has taught us.

Carl also had his humorous and counter-cultural side. I remember him telling me how his lab in the 1960s heard about the rumor that compounds in banana peels were a legal narcotic and how they launched an unofficial research project to isolate these. His verdict was that there was nothing there and neither the peels nor anything in them could get you high — but he wanted to empirically test that. Also, when reading about a supposed “Qi master” who claimed to be able to influence mutation rates with his mind, he invited him to the lab to give a demonstation — which naturally failed to show any effect under controlled conditions — but he wanted to see if the guy could really do it.

Genomics, metagenomics, and evolutionary biology has lost one of its greats — but his legacy lives on.

‘Twas the night before Christmas

‘Twas the night before Christmas, when all through the building
All our creatures were stirring, even our mold;
The dishes were placed in the incubator with prayer,
In hopes that pure growth soon would be there;

The scientists were nestled all close to their screens instead
While swirls of DNA danced in their heads;

My coworker in her labcoat, and I with my pipettor,
Had just settled down for a long overnighter,

When out in the lab there arose such a clatter,
I sprang from my microscope to see what was the matter.
Away to the incubator I flew like a flash,
Tore open the doors then saw what was trash.

When, what to my tired red eyes should appear,
But a bunch of contaminated plates, there goes my career.

Santa Hat - Ho Ho Ho

Brim and ball: Neosartorya fischeri; Hat: Penicillium marneffei; Ho,Ho,Ho: Aspergillus flavus. Image Credit: Stephanie Mounaud / J. Craig Venter Institute.

Last year, still in an isolated fungal room placed far away from others, I made an attempt at this one, but my stocks were contaminated. Something all fungal folks know something about. (Aspergillus is just EVERYWHERE). So with a little luck (let’s face it, with complete luck) I was able to clean things up and told the fungus to be on its best behavior. However, N. fischeri still did not want to play nice with the P. marneffei…so they remained slightly separated.

Fungal Christmas Tree

Star: Talaromyces stipitatus; Tree: Aspergillus nidulans Ornaments: Penicillium marneffei; Trunk: Aspergillus terreus. Image Credit: Stephanie Mounaud / J. Craig Venter Institute.

I hope everyone enjoys my creation, although the credit goes to my jolly ole fungus for being so wonderfully diverse and satisfying my slightly nerdy creative side.

Fungalman

Hat, Eyes, Mouth, Buttons: Aspergillus niger; Arms: Aspergillus nidulans; Nose: Aspergillus terreus with Penicillium marneffei; Body: Neosartorya fischeri. Image Credit: Stephanie Mounaud / J. Craig Venter Institute.

Let us all show the world the true side of fungus and all its amazing potential. Because we all know they can do more than just sit there and look pretty.

JCVI Internship Information for 2013 Is Ready

We are now accepting applications for the 2013 Summer Internship Program.  We are excited to be able to continue to inspire young scientists!  Last year, we received 546 applications.  Of which, thirty-one interns were selected to work in diverse areas.

 

2012 San Diego Summer Interns

2012 San Diego Summer Interns

 

2012 Rockville Summer Interns

2012 Rockville Summer Interns

 

Some of the intern projects were:

  • Isolation and Characterization of Electricity Generating Bacteria
  • Characterizing the Microbial Population of Rabbit GI Tract
  • Cloning Ureaplasma urealyticum: An Odyssey
  • Comparative Genomics of three isolate strains in the actively serpentinizing Cedars Springs
  • Coronavirus: Amplification of the HE and Spike genes in human coronavirus genomes

The 2013 JCVI Internship Program is open to accept spring and summer applications. The application process includes the submission of a resume, essay and transcripts as one PDF file via our online application site. We do not require letters of recommendation.

Information about the 2013 program can be found at http://www.jcvi.org/cms/education/internship-program/

Building a Solid Foundation

The JCVI La Jolla construction site has been busy since earthwork began in 2011.  After grading the site to specified levels, a detailed excavation began to make room for the structural concrete footings, supporting slabs, and underground utilities.  With all of the holes in just the right place, crews began installing steel rebar to provide reinforcement for the large structural footings which will eventually carry the building’s weight.  Once the steel had been placed, it was inspected and filled with concrete.  The largest structural footing was so large that it took 45 concrete trucks to fill the hole!

Concrete wasn’t the only thing being placed under the building.  Of the many sustainable strategies included in the building’s design, two of these systems require large underground water tanks to operate.   The first is a rainwater harvesting system which will collect precipitation from the building roof and site.  Three tanks, totaling 90,000 gallons, collect the water so that it can be treated and then reused for toilets and irrigation.  This system will reduce the building’s potable water demand by 70% annually. Pictured here is a 5,000 gallon fiberglass tank being lowered into position.

5500 gallon rainwater storage tank is lowered into place

To become a net-zero building it is critical that electrical demands are minimized.  The new JCVI La Jolla building is being designed with an ultra-efficient mechanical system to help reduce the building’s energy profile.  At the core of this design are two 25,000 gallon thermal energy storage (TES) tanks.  These tanks will store both warm and cool water which will be used to either heat or cool the building depending on the building’s internal temperatures.  Using water to move heat in and out of the building uses 87% less energy than using air and allows the mechanical systems to operate at much higher energy efficiencies than standard building systems.

Single TES tank arrives by tractor trailer

Two 25,000 gallon TES tanks being installed

Most of the building structure is made with concrete and, after the underground work was complete, McCarthy wasted no time in constructing the vertical support walls.  The walls are architectural concrete because they are designed to complement the other materials such as wood that will make up the building’s exterior.  Using information from the concrete mock-ups (See Moving Dirt at JCVI La Jolla) the team selected a 30% fly-ash concrete mix.  Fly ash is a waste material from coal-fired power plants that lowers the amount of cement needed to make concrete, thus both lowers the energy and carbon dioxide emissions of manufacture and increases the overall volume of recycled content used in building.

Constructing architectural concrete is akin to making art.  Huge forms are built with meticulous detail using special materials, fastening methodologies, and custom routed grooves to create the desired finish.  Those forms are filled with steel rebar, filled with concrete at lightning speed, and then stripped (formwork is removed) at just the right time to produce the final product.  The work is even more challenging given the complex geometries of the building and the performance characteristics of using a 30% fly-ash concrete mix.

 

Erected concrete forms

As of September 2012, the team has almost completed the parking garage and podium deck upon which the building structure will sit. Concrete work will continue through the fall and is expected to complete in early 2013.

 

Mock-up of a single office window at the building’s exterior.

Water testing the window system

JCVI Viral Finishing Pipeline: a Winning Combination of Advanced Sequencing Technologies, Software Development and Automated Data Processing

JCVI viral projects are supported by the NIAID Genomic Sequencing Center for Infectious Disease (GSCID). The viral sequencing and finishing pipeline at JCVI combines next generation sequencing technologies with automated data processing. This allowed us to complete over 1,800 viral genomes in the last 12 months, and almost 8,800 genomes since 2005.

Viral Projects at JCVI

JIRA Viral Sample Tracking Workflow

Our NextGen pipeline, which utilizes SISPA-generated libraries with Roche/454 and Illumina sequencing, enables us to complete a wide variety of viral genomes including challenging samples. Automated assembly pipeline employs CLCbio command-line tools and JCVI cas2consed, a cas to ace assembly format conversion tool. Our complimentary Sanger pipeline software is currently being integrated with the NextGen pipeline. This will improve our data processing and will allow us to use validation software (autoTasker) more efficiently.

Assembly of Repetitive Viral Genomes

Genome Organization of Varicella-Zoster

Assembly of Novel Viral Genomes

CLC Assembly Viewer Representation

Promoter of Bat Genome

Promoter of Bat Genome

During the past year we have found that novel viruses, repetitive genomes, and mixed infection samples could not be easily integrated with our high-throughput assembly pipeline. We have developed an assembly and finishing process that utilizes components of the high-throughput pipeline and combines them with manual reference selection and editing. Using this approach we completed novel adenovirus genomes and mixed-infection avian influenza genomes, and improved assemblies of previously unknown arbovirus genomes. We are currently working on optimizing and automating this new pipeline.

Assembly of Mixed Viral Genomes

Consed Representation of Mixed Viral Sample

Consed Representation of Mixed Viral Sample

Repetitive genomes have long been known to present great challenges during assembly and finishing. We are presenting a new approach to assembly and finishing of repetitive varicella genome that is based on separating it into overlapping PCR amplicons followed by merging sequenced amplicons during assembly.

To streamline our viral pipelines, we have fully integrated them with JCVI’s LIMS and JIRA Workflow Management to create a semi-automated tracking interface that follows the progress of viral samples from acquisition through to NCBI submission. This allows us to process a large volume of samples with limited manual interaction and, at the same time, gives us flexibility to work on challenging and novel genomes.

Acknowledgements

The JCVI Viral Genomics Group is supported by federal funds from the National Institute of Allergy and Infectious Disease, the National Institutes of Health, and the Department of Health and Human Services under contracts no. HHSN272200900007C.

Bat coronavirus project is collaboration with Kathryn Holmes and Sam Dominguez, University of Colorado Medical Center.

The authors would like to thank members of the Viral Genomics and Informatics group at JCVI.

References

Viral genome sequencing by random priming methods. Djikeng A, Halpin R, Kuzmickas R, Depasse J, Feldblyum J, Sengamalay N, Afonso C, Zhang X, Anderson NG, Ghedin E, Spiro DJ. BMC Genomics. 2008 Jan 7;9:5A virus discovery method incorporating DNase treatment and its application to the identification of two bovine parvovirus species.  Allander T, Emerson SU, Engle RE, Purcell RH, Bukh J.

Note

This post is based on a poster by Nadia Fedorova, Danny Katzel, Tim Stockwell, Peter Edworthy, Rebecca Halpin, and David E. Wentworth.

Biowalk of Fame

There is a new “Biowalk of Fame” in Maryland, and our own Craig Venter was one of the first honorees receiving a plaque, which is there for all to see as you stroll through lovely Silver Spring.

Etching of Dr. J. Craig Venter on Biowalk of Fame

Etching of Dr. J. Craig Venter on Biowalk of Fame

Other honorees include Dr. Martin Rodbell and Ben Carson. The event to honor the awardees was on April 22, which also it happens to be Earth Day. Although it rained heavily throughout the event, there were a large number of people in attendance including several local government officials including Council member Valerie Ervin and Chairman Ike Leggett. Dr Martine Rothblatt, CEO of United Therapeutics, emceed the event.

Biowalk of Fame tour sign

Biowalk of Fame tour sign

The idea behind the BioWall and the Biowalk is very innovative. The Wall is a live movie like screen that allows videos from students and the public that relate to science to be continuously aired. A student observing a paramecium under the microscope for example can mail the clip in to United Therapeutics, and it will be available for all to see. The Biowalk also has plaques dedicated to those who have made the most outstanding contributions to the State of Maryland in the sciences – hence Craig.

Dr. J. Craig Venter's plaque on the Biowalk of Fame

Dr. J. Craig Venter's plaque on the Biowalk of Fame

Biowalk of Fame

Biowalk of Fame

The take home message is, if you are wondering through Silver Spring do not be surprised if you see Craig’s name on a plaque on 1040 Spring Street. Congratulations!

Moving dirt at JCVI La Jolla

After celebrating the ground breaking of JCVI La Jolla, McCarthy Building Companies immediately got to work preparing the land for construction.  First the crew set up a work area to house the staff and equipment needed for the project.  The site was cleared and stabilized for construction trailers and a temporary road was built for construction vehicles and equipment.  Water trucks were used to control dust and special shaker plates were installed at the entrance of the site to minimize loose dirt and stones on nearby roads.

With basic infrastructure in place, the team moved next to save three large Torrey Pines growing within the construction zone.  The trees had been identified during the design process and were flagged for relocation to protected areas on the site where they will remain as part of the natural landscape.  Big Trees of California, a firm specializing in relocation of large tree species, began the process of “boxing” these trees in custom built structures 14’ wide by 14’ long and 5’ deep.  Wooden lifting beams were installed underneath to provide connection points for the vertical lift.  A large crane was used to “fly” full grown Torrey Pine trees to their new homes.

Relocating Torrey Pine trees at JCVI La Jolla

With the Torrey Pines now safe, preparations for the building pad began. Several pieces of heavy earth working equipment arrived to begin the dig for the building’s foundation.  However, the crew soon discovered a local soil condition, known as “Lindavista formation,” which proved to be a challenge for even some of the largest machinery.  Fortunately, the team was prepared, and with a few equipment modifications, they reached the designed grading levels required for the excavation in just slightly more time than expected.  The earthen building pad was moisture conditioned, compacted, and surveyed by geotechnical engineers. The first major construction milestone was met!   The team is now focused on installing the concrete foundation and underground utilities.

Caterpillar D-9 at Work

While most of the team was focused on moving dirt, McCarthy’s concrete team began assembling 4’x8’ mock-ups of the architectural concrete that will be used for building’s exterior.  The mock-ups are done to determine the best method for constructing future concrete forms, while simultaneously giving the architects a glimpse at the final appearance of the finished product.  In this first series of concrete mock-ups the team is particularly focused on two important decisions:

1)      How much recycled fly-ash material can be added to the concrete while still maintaining the desired look and strength characteristics? Adding fly-ash to a concrete mix design increases the amount of recycled building material used and can count towards LEED credits. The JCVI building is intended to be one of the few LEED Certified Platinum lab building in the US so every step counts.

2)      How will the concrete forms be constructed to produce the desired finish and house the necessary structural steel elements?


Building forms for architectural concrete mock-ups

Stay tuned for updates on progress of the concrete mock-ups and other design elements of the building.  Also, if you haven’t seen it already, check out the web-cam image here which provides hourly updates from the job site.