Journal of biomolecular techniques : JBT最新文献

This column highlights recently published articles that are of interest to the readership of this publication. We encourage ABRF members to forward information on articles they feel are important and useful to Clive Slaughter, AU-UGA Medical Partnership, 1425 Prince Avenue, Athens GA 30606. Tel; (706) 713-2216: Fax; (706) 713-2221: Email; cslaught@uga.edu or to any member of the editorial board. Article summaries reflect the reviewer's opinions and not necessarily those of the Association.

The cold-shock expression system in Escherichia coli was developed on a manual induction approach using optical density at 600 nm (OD₆₀₀) measurements and isopropyl β-D-1-thiogalactopyranoside (IPTG) addition. In this study, we show that cold-shock expression performs equally well using an autoinduction approach wherein OD₆₀₀ measurements and IPTG addition may be eliminated. We further demonstrate that cold-shock expression with autoinduction can better facilitate high-throughput experiments.

This column highlights recently published articles that are of interest to the readership of this publication. We encourage ABRF members to forward information on articles they feel are important and useful to Clive Slaughter, AU-UGA Medical Partnership, 1425 Prince Avenue, Athens GA 30606. Tel; (706) 713-2216: Fax; (706) 713-2221: Email; cslaught@uga.edu or to any member of the editorial board. Article summaries reflect the reviewer's opinions and not necessarily those of the Association.

Introduction/objective: Academic institutions often struggle to meet the unique professional development needs of shared resource personnel, who require business skills, project and people management expertise, and an active, collaborative network of shared resource colleagues.

Materials and methods: We launched the Vanderbilt Core Exchange professional development and networking program in 2020. The program was intentionally designed with core personnel input and supports faculty and staff from more than 80 shared resources across Vanderbilt University and Vanderbilt University Medical Center. Resources offered include a quarterly seminar series with both virtual and in-person events, a website for accessing professional development materials and session recordings, and a dedicated online discussion group for core personnel networking.

Results: There have been 11 Vanderbilt Core Exchange events to date: 2 in person and 9 virtual. In-person events averaged 35 attendees, and virtual events averaged 45 attendees. Topics included equipment grant writing, marketing, handling difficult conversations, managing different workplace work styles, communication and project management tools, and the importance of self-care. Survey responses collected after each event were highly positive and informed areas of improvement and future event topics.

Discussion: This model of local shared resource professional development serves as a template for institutions who desire to create opportunities for collaboration and community building. With a small coordinating committee of dedicated individuals, an institution-wide professional development and networking program can be successfully established even with limited resources.

Wastewater-based surveillance (WBS) is a noninvasive, epidemiological strategy for assessing the spread of COVID-19 in communities. This strategy was based upon wastewater RNA measurements of the viral target, severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2). The utility of WBS for assessing the spread of COVID-19 has motivated research to measure targets beyond SARS-CoV-2, including pathogens containing DNA. The objective of this study was to establish the necessary steps for isolating DNA from wastewater by modifying a long-standing RNA-specific extraction workflow optimized for SARS-CoV-2 detection. Modifications were made to the sample concentration process and included an evaluation of bead bashing prior to the extraction of either DNA or RNA. Results showed that bead bashing reduced detection of RNA from wastewater but improved recovery of DNA as assessed by quantitative polymerase chain reaction (qPCR). Bead bashing is therefore not recommended for the quantification of RNA viruses using qPCR. Whereas for Mycobacterium bacterial DNA isolation, bead bashing was necessary for improving qPCR quantification. Overall, we recommend 2 separate workflows, one for RNA viruses that does not include bead bashing and one for other microbes that use bead bashing for DNA isolation. The experimentation done here shows that current-standing WBS program methodologies optimized for SARS-CoV-2 need to be modified and reoptimized to allow for alternative pathogens to be readily detected and monitored, expanding its utility as a tool for public health assessment.

This column highlights recently published articles that are of interest to the readership of this publication. We encourage ABRF members to forward information on articles they feel are important and useful to Clive Slaughter, AU-UGA Medical Partnership, 1425 Prince Avenue, Athens GA 30606. Tel; (706) 713-2216: Fax; (706) 713-2221: Email; cslaught@uga.edu or to any member of the editorial board. Article summaries reflect the reviewer's opinions and not necessarily those of the Association.

Core facility laboratories are an essential part of the successful research enterprise of many universities around the world. Core facilities provide state-of-the-art instrumentation and technologies to support research of all faculty, postdocs, and students on a fee-for-service basis. Academic next-generation sequencing cores are typically "full service" facilities, and access to and training on their instrumentation is limited to core staff. To address these limitations, we provided graduate students with technical training at our core facility. We developed a 1-week noncredit-bearing workshop and recruited 6 graduate students (N = 6) as part of a pilot program. The program involved online teaching, classroom-based teaching, and hands-on training in next-generation sequencing library preparation and sequencer operation. A post-participation survey revealed highly positive outcomes in terms of skill development and increased awareness of technologies offered by the core facility. A workshop of this scale could be incorporated into the graduate curriculum and extended to core facilities that focus on other technologies. We believe that introducing formal standardized teaching spearheaded by core facilities would improve the graduate student curriculum and hope that this study can provide guidance on curriculum design for similar workshops.

Flow cytometry is a powerful tool that finds applications in various fields such as immunology, molecular biology, cancer biology, virology, and infectious disease monitoring. A significant portion of the research in these disciplines is supported by flow cytometry shared resource laboratories (SRLs). There are several types of flow cytometers available for use in SRLs, including analyzers, sorters, imaging flow cytometers, and mass cytometers. Each type has different challenges when it comes to maintenance and life expectancy. An independent online survey was conducted to better understand instrument maintenance and turnover in flow cytometry SRLs. Questions regarding instrument uptime (availability), its usage, routine maintenance, and cost associated with it were addressed. The respondents also answered questions pertaining to the frequency of deep cleaning of the instrument and quality control. In addition, the survey queried about the source of funding used to purchase the instruments and possible reasons for a replacement. Presented herein are the results compiled from 146 core facilities that provide a look at the operation within a typical SRL, with the responses reflecting researchers' experiences with handling flow cytometers.

This column highlights recently published articles that are of interest to the readership of this publication. We encourage ABRF members to forward information on articles they feel are important and useful to Clive Slaughter, AU-UGA Medical Partnership, 1425 Prince Avenue, Athens GA 30606. Tel; (706) 713-2216: Fax; (706) 713-2221: Email; cslaught@uga.edu or to any member of the editorial board. Article summaries reflect the reviewer's opinions and not necessarily those of the Association.