Sample Quantification and QC Service Options
Qualitative Analysis by Capillary Electrophoresis
Agilent Bioanalyzer 2100
The Agilent 2100 Bioanalyzer is Agilent's highly successful microfluidics-based platform for the analysis of DNA, RNA, proteins and cells. As the first commercial, analytical instrument based on lab-on-a-chip technology the Agilent 2100 bioanalyzer has proven to be an excellent alternative to messy and labor-intensive gel electrophoresis techniques; delivering fast, automated, high quality digital data instead of the subjective, time-consuming results associated with gels.
Vsit the Agilent 2100 Bioanalyzer website for more information regarding this technology.
For prices and sample volume requirements see the BMGC Quantitation Request Form.
The UMGC offers the following options for Agilent DNA Analysis:
- Agilent High Sensitivity DNA Chip: Low concentrated (5-500 pg/ul) dsDNA samples from 50 - 7000 bp.
- Agilent DNA 1000 Chip: Samples from 0.1-50 ng/ul dsDNA fragments from 25 to 1000 bp.
The UMGC offers the following options for Agilent RNA Analysis:
(you must designate the type of RNA on the QC request form (Eukaryotic Total RNA, Prokaryotic Total RNA, mRNA, Plant RNA, Small RNA)
- Agilent RNA 6000 Nano Chip: For the analysis and quantitation of RNA samples from 25 to 500 ng/µL in concentration.
- Agilent RNA 6000 Pico Chip: For the analysis of low concentrated RNA samples down to 50 pg/µL of total RNA or 250 pg/µL of mRNA.
Caliper GX (New!)
Caliper’s high-throughput platform for nucleic acid separations. This automated microfluidics technology allows for reproducible, high-resolution, electrophoretic separations for 96 samples in under an hour.
Visit the Caliper LabChip® GX website for more information regarding this technology.
For prices and sample volume requirements, see the UMGC Quantitation Request Form. Minimum of 12 samples needed for submission.
The UMGC offers the following Caliper GX DNA Assays:
- DNA 12K: 100-12,000 bp at 1-50 ng/ul
- DNA 5K: 100-5000bp at 1-60 ng/ul
- DNA 1K: 25-1000 bp at 1-60 ng/ul
- DNA High Sensitivity: 50-5000 bp at 25 pg/ul-5ng/ul
The UMGC offers the following Caliper GX RNA Assays:
- RNA: 25-250 ng/ul
- RNA High Sensitivity: 5-50 ng/ul
Note: We recommend cleaning up samples for Caliper GX and Agilent Bioanalyzer Qualitative Analysis. Please elute in water or 10mM Tris-HCL, pH 8.5 for best results. Due to the high sensitivity of the assay, different ions and higher salt concentrations might influence the performance.
The NanoDrop ND-8000 is a full-spectrum (220-750 nm) multi-sample, micro-volume, UV-Vis spectrophotometer that measures 2 ul of sample with high accuracy and reproducibility. It utilizes a patented sample retention technology that employs surface tension alone to hold the sample in place eliminating the need for expensive cuvettes and other devices and allows for clean up in seconds. The ND-1000 also has the capability to measure highly concentrated samples without dilution (50X higher concentration compared to standard specs).
Visit the NanoDrop 8000 website for more information regarding this technology.
PicoGreen and RiboGreen Assays
PicoGreen and RiboGreen are ultrasensitive fluorescent nucleic acid stains for quantitating double-stranded DNA and RNA, respectively, in solution. Unlike the more common technique of measuring absorbance at 260 nm, these assays are relatively unaffected by the presence of common contaminants such as nucleotides, ssDNA, RNA, proteins, and nucleic acid preparation/extraction contaminants. In addition, they are easily adapted for high-throughput quantitation, allowing for the detection of as little as 250 pg/mL DNA and 1ng/mL RNA using a fluorescence microplate reader.
Kapa Library Quantification
Kapa QC is a qPCR-based method for quantitating libraries constructed with Illumina GA adaptors. Unlike other quantitation methods, Kapa QC only detects the amplifiable molecules in a library. Accurate quantitation is very important as this step as the use of inaccurate library concentrations will most likely lead to sub-optimal sequencing capacity.
Visit the Kapa Biosystems website for more information regarding this quantification method
DNA Shearing using the Covaris