This report documents the findings from an onsite audit of the John Seiberling Federal building located in Akron, Ohio. The Federal landlord for this building is the General Services Administration (GSA). The focus of the audit was to identify various no-cost or low-cost energy efficiency opportunities that, once implemented, would reduce in either electrical and gas consumption and increase the operational efficiency of the building. This audit also provided an opportunity to identify potential capital cost projects that should be considered in the to acquire additional energy (electric and gas) and water savings to further increase the operational efficiency of the building.

Since the emerging of second generation sequencing technologies, the evaluation of different sequencing approaches and their assembly strategies for different types of genomes has become an important undertaken. Next generation sequencing technologies dramatically increase sequence throughput while decreasing cost, making them an attractive tool for whole genome shotgun sequencing. To compare different approaches for de-novo whole genome assembly, appropriate tools and a solid understanding of both quantity and quality of the underlying sequence data are crucial. Here, we performed an in-depth analysis of short-read Illumina sequence assembly strategies for bacterial and archaeal genomes. Different types of Illumina libraries as well as different trim parameters and assemblers were evaluated. Results of the comparative analysis and sequencing platforms will be presented. The goal of this analysis is to develop a cost-effective approach for the increased throughput of the generation of high quality microbial genomes.

We have developed two processes with the Biomek FX robot to construct 454 titanium and Illumina libraries in order to meet the increasing library demands. All modifications in the library construction steps were made to enable the adaptation of the entire processes to work with the 96-well plate format. The key modifications include the shearing of DNA with Covaris E210 and the enzymatic reaction cleaning and fragment size selection with SPRI beads and magnetic plate holders. The construction of 96 Titanium libraries takes about 8 hours from sheared DNA to ssDNA recovery. The processing of 96 Illumina libraries takes less time than that of the Titanium library process. Although both processes still require manual transfer of plates from robot to other work stations such as thermocyclers, these robotic processes represent about 12- to 24-folds increase of library capacity comparing to the manual processes. To enable the sequencing of many libraries in parallel, we have also developed sets of molecular barcodes for both library types. The requirements for the 454 library barcodes include 10 bases, 40-60percent GC, no consecutive same base, and no less than 3 bases difference between barcodes. We have used 96 of the resulted 270 barcodes to construct …

The 2009 Participating Load Pilot Customer Impact Evaluation provides evidence that short duration demand response events which cycle off air conditioners for less than thirty minutes in a hot, dry environment do not lead to a significant degradation in the comfort level of residents participating in the program. This was investigated using: (1) Analysis of interval temperature data collected from inside residences of select program participants; and (2) Direct and indirect customer feedback from surveys designed and implemented by Southern California Edison at the conclusion of the program season. There were 100 indoor temperature monitors that were acquired by LBNL for this study that transmitted temperature readings at least once per hour with corresponding timestamps during the program season, June-October, 2009. Recorded temperatures were transferred from the onsite telemetry devices to a mesh network, stored, and then delivered to KEMA for analysis. Following an extensive data quality review, temperature increases during each of the thirty demand response test events were calculated for each device. The results are as follows: (1) Even for tests taking place during outside temperatures in excess of 100 degrees Fahrenheit, over 85 percent of the devices measured less than a 0.5 degree Fahrenheit temperature increase indoors …

Enrichment of emulsion PCR product is the most laborious and pipette-intensive step in the 454 Titanium process, posing the biggest obstacle for production-oriented scale up. The Joint Genome Institute has developed a pair of custom-made robots based on the Microlab Star liquid handling deck manufactured by Hamilton to mediate the complexity and ergonomic demands of the 454 enrichment process. The robot includes a custom built centrifuge, magnetic deck positions, as well as heating and cooling elements. At present processing eight emulsion cup samples in a single 2.5 hour run, these robots are capable of processing up to 24 emulsion cup samples. Sample emulsions are broken using the standard 454 breaking process and transferred from a pair of 50ml conical tubes to a single 2ml tube and loaded on the robot. The robot performs the enrichment protocol and produces beads in 2ml tubes ready for counting. The robot follows the Roche 454 enrichment protocol with slight exceptions to the manner in which it resuspends beads via pipette mixing rather than vortexing and a set number of null bead removal washes. The robotic process is broken down in similar discrete steps: First Melt and Neutralization, Enrichment Primer Annealing, Enrichment Bead Incubation, Null …

This report presents highlights of the research conducted on NERSC computers in a variety of scientific disciplines during the years 2008-2009. It also reports on changes and upgrades to NERSC's systems and services as well as activities of NERSC staff.

This thin, MEMS contact-stress sensor continuously and accurately measures time-varying, solid interface loads over tens of thousands of load cycles. The contact-stress sensor is extremely thin (150 {mu}m) and has a linear output with an accuracy of {+-} 1.5% FSO.

The U. S. Department of Energy’s Intermountain Industrial Assessment Center (IIAC) at the University of Utah has been providing eligible small- and medium-sized manufacturers with no-cost plant assessments since 2001, offering cost-effective recommendations for improvements in the areas of energy efficiency, pollution prevention, and productivity improvement.

This report provides the results of the annual post-closure inspections conducted at the closed Corrective Action Unit (CAU) sites located on the Tonopah Test Range (TTR), Nevada. This report covers calendar year 2009 and includes inspection and repair activities completed at the following seven CAUs: · CAU 400: Bomblet Pit and Five Points Landfill (TTR) · CAU 407: Roller Coaster RadSafe Area (TTR) · CAU 424: Area 3 Landfill Complexes (TTR) · CAU 426: Cactus Spring Waste Trenches (TTR) · CAU 453: Area 9 UXO Landfill (TTR) · CAU 484: Surface Debris, Waste Sites, and Burn Area (TTR) · CAU 487: Thunderwell Site (TTR) The annual post-closure inspections were conducted May 5–6, 2009. All inspections were conducted according to the post-closure plans in the approved Closure Reports. The post-closure inspection plan for each CAU is included in Attachment B, with the exception of CAU 400. CAU 400 does not require post-closure inspections, but inspections of the vegetation and fencing are conducted as a best management practice. The inspection checklists for each site inspection are included in Attachment C, the field notes are included in Attachment D, and the site photographs are included in Attachment E. Vegetation monitoring of CAU 400, …

The High Level Radioactive Waste (HLW) Sludge in Savannah River Site (SRS) waste tanks was produced over a period of over 60 years by neutralizing the acidic waste produced in the F and H Separations Canyons with sodium hydroxide. The HLW slurries have been stored at free hydroxide concentrations above 1 M to minimize the corrosion of the carbon steel waste tanks. Sodium nitrite is periodically added as a corrosion inhibitor. The resulting waste has been subjected to supernate evaporation to minimize the volume of the stored waste. In addition, some of the waste tanks experienced high temperatures so some of the waste has been at elevated temperatures. Because the waste is radioactive, the waste is transforming through the decay of shorter lived radioactive species and the radiation damage that the decay releases. The goal of the Savannah River National Laboratory (SRNL) simulant development program is to develop a method to produce a sludge simulant that matches both the chemical and physical characteristics of the HLW without the time, temperature profile, chemical or radiation exposure of that of the real waste. Several different approaches have been taken historically toward preparing simulated waste slurries. All of the approaches used in the …

Fosmid or BAC end sequencing plays an important role in de novo assembly of large genomes like fungi and plants. However construction and Sanger sequencing of fosmid or BAC libraries are laborious and costly. The current 454 Paired-End (PE) Library and Illumina Jumping Library construction protocols are limited with the gap sizes of approximately 20 kb and 8 kb, respectively. In the attempt to understand the limitations of constructing PE libraries with greater than 30Kb gaps, we have purified 18, 28, 45, and 65Kb sheared DNA fragments from yeast and circularized the ends using the Cre-loxP approach described in the 454 PE Library protocol. With the increasing fragment sizes, we found a general trend of decreasing library quality in several areas. First, redundant reads and reads containing multiple loxP linkers increase when the average fragment size increases. Second, the contamination of short distance pairs (<10Kb) increases as the fragment size increases. Third, chimeric rate increases with the increasing fragment sizes. We have modified several steps to improve the quality of the long span PE libraries. The modification includes (1) the use of special PFGE program to reduce small fragment contamination; (2) the increase of DNA samples in the circularization step …