BioMicroCenter:Advanced analytical Fragment analyzer
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Purchased in July of 2012, the Advanced Analytical Fragment Analyzer (FA) is a highly efficient and accurate fluorescence-based electrophoresis instrument used to size and quantify high sensitivity DNA and both nano (conc. >5ng/ul) and pico RNA (conc. 50 pg/ul-5ng/ul). It is faster and more cost effective than the BioAnalyzer2100, and can handle unsupervised runs of up to 96 samples. With a failure rate of less than one percent, the machine’s efficiency allows the BMC to analyze most samples within 24 hours of submission.
In 2017, the FA was upgraded to handle 48 samples in a batch. Each run takes ~60 minutes and 2 μL of sample with a cost of $8 per sample. Users may submit any volume of sample with the expectation that if there is less than 2 μL, the samples will be diluted. To save money and submit a larger quantity of samples, users may come into the BMC and obtain a special 96 well plate or plates. For mass submission, 2 μL must be put into each sample well row by row. There is a way to submit rows in iLabs, but please remember to come and get special plates from the BMC to do mass submission.
High Sensitivity DNA
The FA analyzes DNA 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. 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. For a more in-depth description see the link below:
The FA processes DNA fragments of 50-5000bp at a wide range of concentrations:
Longer fragments and lower inputs can be handled using the Femto Pulse
Nano and Pico RNA
The Advanced analytical 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. It is capable of analyzing both pico and nano RNA.
The Advanced analytical 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 50pg/ul-5ng/ul for pico RNA or 5ng/ul-500ng/ul for Nano RNA.
lower inputs can be handled using the Femto Pulse
The Advanced Analytical is capable of identifying problems such as hyperamplification and “primer-dimer”.
Platform Comparison with BioAnalyzer 2100
|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||60' for 12 samples|
|Prep/Loading Time||>40' for 10 samples||<20' for 48 samples||<5' for 12 samples|
|Minimum Conc.||~0.02nM (DNA)||~0.06nM (DNA)||~0.5 pM|
|Cost Per Sample||$15||$8||TBD|
|Assays||Many||High Sens. DNA, Nano RNA, Pico RNA||ultra sensitive DNA and RNA, long insert DNA|