BioMicroCenter:FemtoPulse
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The FEMTO Pulse (FP) is an instrument that provides a remarkable upgrade over the existing AATI Fragment Analyzer and Agilent BioAnalyzer technologies that have been available for many years. While still an instrument to analyze the lengths of nucleic acids, the FP adds a pulse field generator to allow the separation and characterization of significantly longer molecules, out to 200,000 base pairs. Additionally, the FP has significantly improved optics that allow approximately two orders of magnitude of additional sensitivity. Where the AATI Fragment Analyzer bottoms out a sensitivity of 2-3pg/ul, the FP has nearly 100x improved sensitivity which may enable this instrument to directly observe molecular reactions without the need for amplification or modification with radioactive nucleotides.
The FP runs 12 samples at a time using 2ul of material; for pricing, please see our pricing page. To save money when submitting a higher quantity of samples, users can bring a preloaded plate with 2uL of sample, organized by row; these plates will be run as submitted, without dilution. Each run takes ~90 minutes. It is *not* a replacement for the Fragment Analyzer and is optimized for very low inputs or very long samples.
The FEMTO Pulse was donated to the BioMicro Center by MIT CEHS.
Ultra High Sensitivity/Genomic DNA
The FP analyzes DNA using pulse-field capillary electrophoresis. A tiny amount of sample is injected through a capillary with gel, and across a fluorescence detection window using a voltage gradient. DNA intercalating dye within the gel makes the DNA detectable by a camera lens, which is able to read the fluorescence levels of the fragments through the window, and accurately give the size and concentrations of the sample. Unlike the Fragment Analyzer, the pulse-field generator of the FP enables sizing of much longer DNA fragments, up to 200kbp in length.
The FA processes DNA fragments of 50bp-200kbp at a wide range of concentrations:
0.1pg/μL-100pg/μL (Fragments)
2.5pg/μL-500pg/μL (Smear)
0.3pg/μL-30pg/μL (gDNA Fragment)
5pg/μL-500pg/μL (gDNA Smear)
Higher inputs can be handled using the Fragment Analyzer.
Ultra High Sensitivity RNA
The FP analyzes RNA also using capillary electrophoresis. A tiny amount of sample is injected through a capillary with gel, and across a fluorescence detection window using a voltage gradient. RNA intercalating dye within the gel makes the RNA detectable by a camera lens, which is able to read the fluorescence levels of the fragments through the window, and accurately give the size and concentrations of the sample.
The FP provides an RNA quality number (RQN) from 1-10 on samples of nano or pico RNA. It is capable of analyzing samples at concentrations of 2.5-500pg/μL.
Higher inputs can be handled using the Fragment Analyzer.
Platform Comparison
| BioAnalyzer2100 | Fragment Analyzer | FEMTO Pulse | |
|---|---|---|---|
| Volume per sample | 1 μL | 2 μL | 2 μL |
| Time per run | 45' for 10 samples | 60' for 48 samples | 90' for 12 samples |
| Prep/Loading Time | >40' for 10 samples | <20' for 48 samples | <5' for 12 samples |
| Failure Rate | 15-30% | <1% | unknown |
| Minimum Conc. | ~0.02nM (DNA) | ~0.06nM (DNA) | ~0.5 pM |
| Assays | Many | High Sens. DNA, Nano RNA, Pico RNA | ultra sensitive DNA and RNA, long insert DNA |
