Sample Preparation Equipment/Instruments

For access or consultation: Dr. Kim DeWeerd 591-8831 or

The Molecular Core also has an Ohaus triple beam balance, Mettler Toledo AB204-S/Fact analytical balance, Denver Instruments APX-153 top loading balance, Mettler Toledo Five easy pH meter and a Beta Star Autoclave. The Molecular Core also has one Baker Company Sterilgard III Advance laminar flow hood, one Coy Lab Products Type B anaerobic chamber, one Diagenode Disruptor 300, one Branson sonifier, one Kinematica Polytron PT1330D homogenizer, one BioRad Genepulser Xcell Electroporator and one Barnstead MiVac Duo concentrator/evaporator.

Sample preparation is a critical component to any experiment. Basic procedures like using nanopure water, weighing chemicals, measuring pH of buffers and medium and sterilizing glassware, utensils, buffer are essential to all experiments. Sample preparation includes manipulating cells, tissues or biological materials in an appropriate environment, breaking open cells or homogenizing cellular material to isolate biological components, concentrating and drying samples and even moving DNA into cells. The Molecular Core facility has instruments that are designed to facilitate sample preparation.

Specific Description:
The laminar flow hood is designed to direct airflow around a surface where cell and tissue manipulation occurs to minimize contamination. The atmosphere is continuously filtered with HEPA filters and ultraviolet light illumination keeps the surface sterile while not in use.

The anaerobic chamber maintains an atmosphere of 97% N2 and 3% H2 through catalytic removal of O2 gas with a palladium catalyst and using an airlock to bring samples or materials in from outside of the chamber. This ensures an anoxic environment from which to prepare samples or microorganisms that are sensitive to oxygen.

The Branson SLPe sonifier can be used to facilitate solubilization of certain chemicals in aqueous buffers or to break the cell walls of microorganisms. The Branson sonifier can be run in pulse or continuous mode with a 40 KHz ultrasonic frequency and a maximum power output of 150 Watts.

The Diagenode Disruptor can also do these tasks as well as shear DNA to facilitate DNA methylation, chromatin immunoprecipitation and high throughput DNA sequencing. The Diagenode uses a sonicating, refrigerated water bath and motor that rotates samples in the bath at a fairly constant temperature inside a sound dampening box.

The Kinematica Polytron is used to homogenize insoluble tissues and cells to aid in releasing desired cellular components. the Kinematica uses a 24 Volt, 100 Watt motor and digital speed control of 2,000 to 30,000 rpm to produce aggregate tip speeds up to 14 m/s to produce the smallest particle possible.

The electroporator is used to move plasmid or other DNA into cells. The Gene Pulser Xcell provides electroporation of all cell types with preprogramed parameters for twelve mammalian cell types, 7 bacterial species and five fungal species. The electroporator can store up to 144 different electroporation programs and can deliver a maximum of 3,000 V.

The Barnstead MiVac is used to remove aqueous solutions in order to concentrate samples or change solvents. The MiVac Duo has a centrifuge for spinning 96 well plates at 1000 rpm with a programmable temperature range of ambient to 99°C and programmable time of 0 to 99 minutes. It also contains a -50°C condensation trap and a vacuum that achieves 10 mbar.

For More Information:
Gene PulserXcell Brochure
Branson Sonifier Manual
Diagenode Disruptor Manual

Instrument Photographs:
Click to enlarge