Quantity control of nucleic acids with a fluorospectrophotometer Nanodrop 8000 (Thermo Scientific)
As a part of the isolation service, we will measure the quantity and quality of you NA with Nanodrop 8000 spectrophotometer, which can realiable concentrations from 2.5 to 3,700 ng/µL dsDNA.
A measured values and their optimal ranges:
- DNA a RNA (260nm) > 10 ng/ul
- A good quality DNA sample should have a A260/A280 ratio of 1.7-2.0 and an A260/A230 ratio of greater than 1.5
- A good quality RNA sample should have a A260/A280 ratio of 1.8-2.0 and an A260/A230 ratio of 1.8 – 2.2
ADDITIONAL RESOURCES:
How to interpret RNA peaks: http://www.u.arizona.edu/~gwatts/azcc/InterpretingSpec.pdf
What contamination of your NA looks like: http://www.nanodrop.com/Library/T009-NanoDrop%201000-&-NanoDrop%208000-Nucleic-Acid-Purity-Ratios.pdf
Importance of 260/230 and 260/280 ratios: http://www.nanodrop.com/Library/T042-NanoDrop-Spectrophotometers-Nucleic-Acid-Purity-Ratios.pdf
Troubleshooting for RNA isolation: http://bitesizebio.com/2345/troubleshooting-rna-isolation/
Quantity control of nucleic acids with a fluorospektorphotometer Quibit 2.0 (Life Technologies)
We offer quantification of nucleic acids using The Qubit® Fluorometer that utilizes fluorescent dyes that are specific to the target of interest. Qubit® assay kits fluoresce only when bound to the selected molecule—DNA, RNA, or protein—in your sample, even at low concentrations. It relies on reading two standards for calibration every time that you make measurements. From 1 to 20 µl of DNA sample is required, with 2 µl being the amount typically used by the lab. Researchers can use instrument after training or we can isolate your cells for you in form of a service.
Advantages:
- Ideal for measurement of low concentrations NA below 5 ng/ul because UV spectrophotometry often does not have the sensitivity to accurately measure low concentrations of DNA and RNA.
- The effective range covers a sample concentration range of 10 pg/μL to 1 μg/μL DNA.
- Qubit® RNA assays are designed to be specific and accurate for RNA even in the presence of a 1:1 mixture of RNA and DNA.
- The Qubit® Fluorometer supports assays for high-sensitivity or broad-range dsDNA and RNA quantitation. Assays for oligos, ssDNA, protein, and microRNA quantification are also available.
ADDITIONAL RESOURCES:
https://www.thermofisher.com/content/dam/LifeTech/migration/en/filelibrary/cell-tissue-analysis/qubit-all-file-types.par.0519.file.dat/qubit-2-fluorometer-user-manual.pdf
https://www.thermofisher.com/cz/en/home/industrial/spectroscopy-elemental-isotope-analysis/molecular-spectroscopy/fluorometers/qubit/qubit-assays.html
https://www.youtube.com/watch?v=RRKZN–7jqg
COST OF A SERVICE: 13 CZK without VAT per a sample
Integrity control of nucleic acids with an Experion Automated Electrophoresis Station (Bio-Rad)
We offer integrity control of nucleic acids using Experion Automated Electrophoresis Station that replaces regular gel analysis and offers higher sensitivity. The instrument combines quantitation and quality assessment in a single step. From 1.5 to 2.5 µl of NA sample is required. Kit for measuring RNA is available at CF. 12 samples of RNA or 11 samples of DNA can be measured in one chip, respectively. Researchers can use instrument after training or we can measure integrity your samples for you in form of a service.
Integrity of RNA: The RQI (RNA Quality Indicator) method returns a number between 10 (intact RNA) and 1 (highly degraded RNA) for each eukaryotic RNA sample run on an Experion RNA StdSens or HighSens analysis chip. The RQI measures RNA integrity by comparing the electropherogram of RNA samples to a series of standardized degraded RNA samples.
DNA analysis: The Experion DNA 1K and DNA 12K analysis kits allow analysis of DNA samples with size ranges of 15–1,500 bp and 50–12,000 bp, respectively. These DNA assays provide high sensitivity and excellent resolution (down to 5 bp) over a broad dynamic range. Consuming only 1 µl of sample for each analysis, the Experion automated system can analyze 1–11 samples in less than 40 min. These assays are recommended for analysis of restriction digests, amplified DNA, microsatellites, and AFLPs.
Protein analysis: The Pro260 kit offers the ability to analyze one to ten protein samples (10–260 kD) in approximately 30 min.
ADDITIONAL RESOURCES:
http://www.biorad.com/webroot/web/pdf/lsr/literature/BULLETIN_5761B.pdf
http://www.bio-rad.com/webroot/web/pdf/lsr/literature/Bulletin_5868A.pdf
http://www.bio-rad.com/webroot/web/pdf/lsr/literature/Bulletin_5909.pdf
INTEGRITY CONTROL OF NUCLEIC ACIDS WITH A BIOANALYZER 2100 (AGILENT)
We offer integrity control of nucleic acids using Agilent 2100 BioAnalyzer that replaces regular gel analysis and offers higher sensitivity. The instrument combines quantitation and quality assessment in a single step. From 1.5 to 2.5 µl of NA sample is required. Kit for measuring RNA is available at CF. 12 samples of RNA or 11 samples of DNA can be measured in one chip, respectively. Researchers can use instrument after training or we can measure your samples for you in form of a service.
Integrity of RNA:
The RIN (RNA Integrity Number) algorithm calculates RNA integrity using a Bayesian learning network that takes into consideration 8 features from the electrophoretic trace, including total RNA ratio (rRNA peaks to total area of the electropherograms), height of the 18S rRNA peak, ratio of the fast migrating RNAs to the total area of the electropherograms, and height of the lower marker (Imbeaud et al. 2005). The output RIN is a number between 1 (completely degraded RNA) and 10 (intact RNA).
- Agilent RNA 6000 Nano Kit for the analysis and quantification of total and mRNA samples of 25 to 500 ng/µL.
- Agilent RNA 6000 Pico Kit for the analysis and quantification of total and mRNA samples of 50 – 5000 pg/µL.
- Agilent Small RNA kit for the analysis and quantification of small RNA samples of 6 and 150 nt in size and 50 to 2000 pg/µL.
Analysis of DNA:
The DNA kits are ideal for automated sizing and quantitation of PCR fragments, restriction digests or fragmented DNA.
The DNA 1000, 7500 and 12000 kits offer automated analysis of PCR fragments, restriction digests and next-generation sequencing (NGS) samples. Typical application areas are: amplicon QC in forensic DNA studies, gene expression analysis by RT-PCR, DNA QC for NGS, or detection of genetically modified organisms.
The High Sensitivity DNA kit allows analysis of fragmented DNA or DNA libraries sith increased sensitivity – down to 5 pg/µL. Analysis of complex DNA samples – down to 100 pg/µL for fragmented DNA or DNA libraries.
ADDITIONAL RESOURCES:
RNA Integrity Database: http://agilent.cnpg.com/Video/flatFiles/194/
http://www.agilent.com/cs/library/applications/5989-1165EN.pdf
Integrity control of nucleic acids with a Fragment analyzer (Advanced Analytical)
We offer measurement of integrity control of your nucleic acids using Fragment Analyzer that is powerful automatic capillary electrophoresis. It covers the full spectrum of DNA, gDNA and RNA quality control. Fragment Analyzer™ does everything from high-resolution analysis to fast DNA separations, across the widest separation range (2 – 40 000 bp). Detection starts from 5 pg/μL. For 12 samples.
Quantitative Analysis Kits: http://aati-us.com/product/fragment-analyzer/reagents
- DNA/NGS Fragment Analysis
- Large Fragment Analysis
- Genomic DNA Analysis
- RNA Analysis
- Qualitative/Sizing Analysis Kits
ADDITIONAL RESOURCES: http://aati-us.com/sites/default/files/FA%20Brochure.pdf
