Quality checking/assessment of DNA and RNA
Quality analyses of DNA and RNA on Agilent BioAnalyzer
The Agilent 2100 BioAnalyzer system provides sizing, quantitation and quality control of DNA and RNA on a single platform, providing high quality digital data.
- Ready-to-use assays and pre-packaged reagent kits
- Minimal sample consumption (1µL)
- Digital data for convenient analysis, archiving and storage
- Contamination-free switch of methods
Check the Agilent BioAnalyzer website for more details. For instance about salt requirements.
Many different applications are available for the 2100 BioAnalyzer but we only keep a few different chips and reagents in stock (see below). Bring your samples in individual 1.5-ml Eppendorf tubes. Although just 1µl is needed of each sample please provide at least 3µl and whatever is left you get it back if samples are precious. Reagents for the following applications are usually kept in stock:
- Agilent DNA 1000 Assay (12 samples/chip, 25–1000bp, 0.1-50ng/µl)
- Agilent High Sensitivity DNA Assay (11 samples/chip, 50-7000bp, 5-500pg/µl)
- Agilent RNA 6000 Nano Assay (12 samples/chip, 5-500ng/µl)
- Agilent RNA 6000 Pico (High Sensitivity) Assay (11 samples/chip, 50-5000pg/µl)
For any other application, check the BioAnalyzer web site (see above) and if any other protocol is of interest please make an inquiry.
For all assays listed, we charge 500 SEK/chip, except for the Agilent High Sensitivity DNA Assay for which we charge 600 SEK/chip. On top of that, we add a 30% service charge (as of 2019-04) on the total to cover for other consumables and service contracts. For all users “overhead” charges are additionally applied. For users outside Lund University we also need to charge for labor on an hourly basis.
Sample form – BioAnalyzer Assessments
A Sample Form (see BioanalyzerForm; .xlsx; 37 kB) for analysis on Agilent BioAnalyzer is required providing information about the client, invoicing information and samples provided. Please label your tube(s) numerically in accordance. If more than one full-chip run is requested, one Sample Form for each chip should be provided.
Quantitative fluorometric analyses of DNA and RNA (PicoGreen/RiboGreen)
Using the PicoGreen dsDNA quantitation assay, you can selectively detect as little as 25 pg/ml of dsDNA in the presence of ssDNA, RNA, and free nucleotides. The assay is linear over three orders of magnitude and has little sequence dependence, allowing you to accurately measure DNA from many sources, including genomic DNA, viral DNA, miniprep DNA, or PCR amplification products. Using the RiboGreen total RNA quantitation assay, you can selectively detect single-stranded RNA down to 1ng/ml of RNA. The reagent is one of the most sensitive dyes for the quantitation of RNA in solution. The RiboGreen reagent also binds to DNA to some extent, and pretreatment of mixed samples with DNase can be used to generate an RNA-selective assay. For both assay types that are run on QuantiFlour ST fluorometer, there is a high- or low-standard range that is chosen and indicated in the sample form.
For both PicoGreen and RiboGreen measurements, there is a series of eight standard points included, and we then charge 12 SEK/sample + 96SEK for the standard. On top of that, we add a 30% service charge (as of 2019-04) on the total to cover for other consumables and service contracts. For all users “overhead” charges are additionally applied. For users outside Lund University we also need to charge for labor on an hourly basis.
Sample form – PicoGreen/RiboGreen measurements
A Sample Form (see FluorometerForm; .xlsx; 31 kB) for quantitative analysis using PicoGreen or RiboGreen is required providing information about the client, invoicing information and samples provided. Please label your tube(s) numerically in accordance.
The sequencing Facility is situated at Department of Biology, in the Ecology Building, Sölvegatan 37 on Floor 2 (staircase A).
For inquiries about Illumina MiSeq sequencing, analyses on Agilent BioAnalyzer and/or fragmentation of DNA/RNA on Covaris, contact:
Telephone: +46 46-222 45 49
Mobile: +46 709-57 18 04
E-mail: Tomas [dot] Johansson [at] biol [dot] lu [dot] se
For inquiries about ABI Sanger Sequencing and Fragment Analysis contact:
Telephone: +46 46-222 08 70
Mobile: +46 72-203 33 56
E-mail: Anna [dot] Sterngren [at] biol [dot] lu [dot] se
The facility acknowledges generous financial support during the years from the Knut and Alice Wallenberg Foundation (SWEGENE programme), the Crafoord Foundation and the Faculty of Science, Lund University.