About

Molecular biology approaches are utilized to understand the mechanisms underlying respiratory diseases caused by respiratory pathogens. The overall objective of the Molecular Biology Core (MBC) is to provide centralized services and technical expertise in molecular biology for Phase III CoBRE and non-CoBRE investigators. Under the Phase I CoBRE award, the MBC was established to provide services involving molecular biology technologies, including RNAi gene silencing, transgene expression, mutagenesis, gene and pathway reporter assays, real-time PCR, and DNA microarray analyses for gene profiling. State-of-the-art technologies and other services based on the needs of the CoBRE investigators have been continuously developed and added to the Core since its inception. In Phase II, the MBC was expanded to include new techniques, including CRISPR/Cas9 technology and bulk RNA sequencing, and to offer reagents for respiratory and infectious disease research, including cell lines, primary lung cells, and respiratory viruses and bacteria. This Phase III application describes additional cutting-edge technologies and valuable reagents that will be offered to Center investigators, including single-cell RNA sequencing, spatial gene profiling, genome-wide CRISPR screening, human lung tissue models (lung organoids) and clinical samples from respiratory infectious disease patients. The specific aims of the MBC are to provide services and support for 1) functional studies using RNAi and CRISPR/Cas9, transgene overexpression, mutagenesis and deletion, lentiviral, adenoviral and nonviral vectors for gene transfer, and stable knockout and transgenic cell lines, 2) genome-wide gene profiling using bulk RNA sequencing, single-cell RNA sequencing, and spatial gene profiling, 3) genome-wide CRISPR gene activation, knockout and interference screens, 4) molecular assays including reporter assays, real-time PCR and droplet digital PCR, 5) reagents for respiratory and infectious disease research including respiratory viruses and bacteria, and reporter viruses, cell lines, primary lung cells, human lung organoids and clinical samples from respiratory infectious disease patients, and 6) advice, consultation and training on various aspects of molecular biology, developing or adapting new methods as needed and disseminating core resources and technologies.

OCRID Core Facilities Service Request Form

Please use the online form linked below to request a service from our core facility. Thank you!

Molecular Biology and Genomics Core Service Request Form

OCRID Core Facilities User Satisfaction Survey

OCRID core facilities are committed to providing top quality services and technical expertise in respiratory and infectious disease for center investigators as well as non-center investigators. We encourage you to fill out our quick 10 question survey so your answers, comments and suggestions can help us improve our services. 

Core Facilities User Satisfaction Survey

• People

  Core Director 

  Lin Liu, Ph.D, CVHS, OSU
  Regents Professor of Physiological Sciences
  Lundberg-Kienlen Endowed Chair in Biomedical Research
  Director, Oklahoma Center for Respiratory and Infectious Diseases
  Center for Veterinary Health Sciences
  Oklahoma State University-Stillwater 

  Contact Information:

  E-mail: lin.liu@okstate.edu
  Phone: (405) 744-4526
  Office: 210 McElroy Hall, Stillwater, OK 74078

  Biosketch: Dr. Liu

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  Core Personnel

  Chaoqun Huang, M.D., M.S., Ph.D.
  Associate Director and Associate Research Professor
  Office: Rm 209A, McElroy Hall
  Email: chaoqh@okstate.edu

   

   

   

   

   

   

   

  Yurong Liang, Ph.D.
  Core Manager
  Office: Rm 206 McElroy Hall
  E-mail: yurong.liang@okstate.edu
  Phone: 405-744-1686

   

   

• Standard Operating Protocol 

  MBC: Standard Operating Protocol (SOP)

  • Initiation of a project by PI
  • Consultation meeting
    • Experimental design
    • Number of samples
    • Timelines etc.
  • Assigned to MBC personnel
  • A report to PI
  • A meeting for result interpretation and problem solving.
• Services

  Services provided by the Molecular Biology Core

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  Functional studies

  • Loss-of-function: RNA interference (RNAi) and CRISPR/Cas9
  • Gain-of-function: Transgene overexpression
  • Mutagenesis/deletion
  • Adenoviral, lentiviral or nonviral vectors for gene transfer in vitro and in vivo
  • Generation of knockout and transgenic cells

  Genome-wide profiling (mRNA, microRNA and lncRNA)

  • Bulk RNA sequencing
  • Single-cell RNA sequencing
  • Spatial gene profiling (GeoMx)

  Genome-wide screens

  • CRISPR activation screens
  • CRISPR knockout screens
  • CRISPR interference screens

  Molecular Assays

  • Reporter assays
  • Real-time PCR and droplet digital PCR

  Reagents

  • • Respiratory viruses and bacteria
    • Reporter viruses (IAV, RSV and SARS-CoV-2)
    • Continuous cell lines
    • Primary lung cells
    • Human lung tissue models (lung organoids)
    • Clinical samples from respiratory infectious disease patients (nasal and airway epithelial cells, bronchoalveolar lavage cells, and peripheral blood mononuclear cells)

  Consultation and training on various aspects of molecular biology

   

  Develop or adopt new methods as needed

   

• Equipment 

  	X10 Genomics Chromium X  Chromium X offers the high-throughput single cell RNA sequencing analysis
  	QX200 Droplet Digital PCR System The QX200 Droplet Digital PCR (ddPCR) System provides absolute quantification of target DNA or RNA molecules for EvaGreen or probe-based digital PCR applications.
  	Nikon Ti2-E Motorized microscope The ECLIPSE Ti2 inverted microscope delivers an unparalleled 25mm field of view (FOV) that revolutionizes the way you see. With this incredible FOV, the Ti2 maximizes the sensor area of large-format CMOS cameras without making compromises, and significantly improves data throughput.
  	Nanosight NS300 This versatile instrument offers the major benefit of minimum user set-up,  enhanced concentration measurements via advanced algorithms and fluorescence detection capability providing truly holistic analysis.
  	Thermal Cycler Common features of Bio-Rad's thermal cyclers include Peltier effect technology for precise temperature control, and thermal gradient to easily optimize PCR assays in a single run.
  	Real-Time PCR CFX Real-Time PCR Detection Systems are powerful and flexible instruments, featuring 2–5 color multiplexing, advanced optical technology, and precise temperature control with thermal gradients.
  	Nanodrop This compact instrument delivers full spectrum UV-Vis analysis and is ideal for rapid nucleic acid and protein quantification.
  	Incu-ShakerTM This instrument's outstanding temperature accuracy and uniformity are supported by proprietary heat distribution technology that ensures that temperatures are precisely maintained throughout the chamber.
  	Microarray Scanner  The instrument has three lasers (488/514nm, 543nm, 633nm) for maximum flexibility and is capable of scanning resolutions of 100mm to 5mm.
  	Amaxa Nucleofector System Nucleofector® Technology offers transfection with high efficiency, low cell mortality and no detrimental physiological effect upon cells,the unique pulses deliver substrate not only to the cytoplasm but also directly to the nucleus.
  	Fluostar Optima Microplate Reader (BMG Biotech) By integrating a high intensity xenon flash lamp with sophisticated light guide technology the FLUOstar OPTIMA covers the full wavelength range 230 to 900 nm for both excitation and emission. The FLUOstar OPTIMA can read all plate formats from 6- up to 384-well plates in all four measurement modes.

   

• Forms

  Sample Submission Forms 

  • Real Time PCR
  • RNA Sequencing 
  • Vector Construction 
  • Other Requests