The FEMTO Pulse is a versatile instrument with various applications across many research disciplines. Capable of separating large DNA fragments and smears, BAC clones, total RNA, and small RNA with unmatched speed, the list of potential applications of the FEMTO Pulse goes on and on. The researchers’ imagination is the only limiting factor for the FEMTO Pulse.
|Maximum Sample Throughput||12 samples per run|
|Maximum Unattended Sample Capacity||288 samples|
|Minimum Sample Volume||20 µL of liquid for injection; as little as 1-2 µL of actual sample required|
|Quantitative Kits||NGS Library Preparations for Illumina and PacBio, Genomic DNA, cfDNA, Total RNA, mRNA, ribodepleted RNA, microRNA, and DNA Fragments|
|Sizing Accuracy||Typically 5% or better|
|Detection Sensitivity||As low as 5 fg/µL for fragments and 200 fg/µL for a smear|
|Light Source||2X 700 mA, 10 W LED, 460 nm excitation wavelength|
|Detector||High sensitivity CCD; 500-600 nm emission wavelength|
|Software||FEMTO Pulse Operational Software, PROSize® 3.0 Data Analysis Software|
|Data Export Format||CSV, PDF, flexible numerical or binary output options|
|Environmental Conditions||Indoor use, normal laboratory environment 20-23°C|
|Relative Humidity Range||<80% (non-condensing)|
|Electrical||100-200 VAC; 50-60 Hz; 15 A (alternate configurations available)|
|Instrument Dimensions||Fully configured 61 cm H x 38 cm W x 51 cm D (24 x 14 x 20)|
|Instrument Weight||40 Kg (88 lbs)|
The FEMTO Pulse Automated Pulsed-Field CE Instrument is a novel tool for genomics and molecular biology research. A remarkably flexible instrument, the power of CE is combined with pulsed-field electrophoresis and an enhanced optical detection system. Pulsed-field capillary electrophoresis (PFCE) supplies researchers with the means to quickly and efficiently separate large DNA fragments and smears. A methodically designed optical detection platform gives the FEMTO Pulse the sensitivity to detect DNA and RNA samples down to the single femtogram level. The pulsed-field power supply is versatile, enabling both PFCE and standard capillary electrophoresis.
In combination these factors produce an automated, high-throughput workflow for the analysis of a wide variety of DNA and RNA samples
PFCE with the FEMTO Pulse provides a streamlined and easy to use alternative to pulsed-field gel electrophoresis (PFGE). The FEMTO Pulse is able to separate smears in about one hour and DNA fragments in less than 2 hours, while PFGE separation time can extend beyond 20 hours.
The FEMTO Pulse Automated Pulsed-Field CE Instrument has innumerable applications and supports researcher innovation. With sensitivity in the femtogram range and sizing through 200,000 bp, new measurements can be made and novel applications developed. The FEMTO Pulse is poised to have a significant impact on various applications including:
The FEMTO Pulse does not require amplification with fluorescently-labeled probes to achieve a remarkable level of detection sensitivity, instead a sensitive intercalating dye is used to detect DNA and RNA. This strategy allows the FEMTO Pulse to analyze nucleic acids without prior knowledge of the samples; in turn broadening the analytic scope of the instrument. What could you do with an instrument as sensitive and powerful as the FEMTO Pulse?
Agarose gel electrophoresis lacks the capability to resolve DNA fragments and smears starting around 20 Kb, in a phenomena termed, "limiting mobility". Functionally, limiting mobility results in the co-migration of large DNA fragments in excess of the size threshold as they are unable to migrate in a size dependent manner. Pulsed-field gel electrophoresis (PFGE) ameliorates these issues by alternating the direction of the electric field. This technique uses an agarose gel to separate fragments up to 10 Mb in size. While PFGE is a powerful technique, enabling the visual analysis of large DNA fragments and smears – including intact genomic DNA – it is time consuming, requiring days of preparation time, significant amounts of genomic DNA, and electrophoresis times between 15 and 24 hours. The FEMTO Pulse Automated Pulsed-Field CE Instrument accelerates this workflow significantly, decreasing preparation time and completing pulsed-field capillary electrophoresis (PFCE) runs in approximately 1 hour.
The FEMTO Pulse is equipped with a pulsed-field power supply, enabling pulsed-field capillary electrophoresis (PFCE). The power supply is additionally capable of supplying constant voltage for the separation of nucleic acid fragments and smears that do not require PFCE. To separate large nucleic acid smears and fragments, the FEMTO Pulse periodically changes the direction of the electric field by 180° in a specific type of pulsed-field electrophoresis called field-inversion electrophoresis. Alternating the electric field in such a manner overcomes limiting mobility, forcing the size dependent separation of large DNA fragments and smears.
An optimized optical system facilitates the detection of nucleic acid samples down to 5 fg/µL. A general overview of how nucleic acids are detected on the FEMTO Pulse is as follows; an LED light source (470 nm) passes through the capillaries in the detection window, exciting the dye intercalated into the DNA or RNA molecules. The emitted fluorescence then passes through a camera lens and specific band pass filters and spatially imaged onto a CCD detector providing results in real-time.
A custom 12 Capillary Ultra-Short Array is employed with the FEMTO Pulse Automated Pulsed-Field CE Instrument. With an effective length of 22 cm and an overall length of 40 cm, the 12 Capillary Ultra-Short Array is shorter than the capillary arrays used with either the Fragment Analyzer Automated CE System or the Fragment Analyzer INFINITY Automated CE System. The shorter length keeps Pulsed-Field Capillary Electrophoresis (PFCE) separation times low without compromising nucleic acid fragment and smear resolution. In addition to a shorter effective length and overall length, the detection window on the 12 Capillary Ultra-Short Array has been modified to reduce crosstalk between capillaries and facilitate enhanced detection sensitivity.