DNA and RNA Extraction FAQs

Our leaf sampling kit (KBS-9370-110) requires 6 mm leaf punches to be collected from each sample and placed into the wells of 96-well plates. As a rough guide, we typically extract 1 µg of DNA per leaf punch although this obviously varies considerably according to species, age of tissue, etc. It is important not to sample too many punches per well of the plate as this can make homogenisation of the tissue very difficult. Conversely, if you know from experience that you extract relatively low quantities of DNA from your tissue, you may want to provide a couple of extra punches to allow for this.

For each SNP we require 5-50 ng of DNA per sample, depending on genome size. For example, if you are planning to run 10 SNP assays across your samples, you will need to send sufficient material for extraction of between 50 and 500 ng DNA. If there is the possibility that you will run further assays on these samples in the future, ensure that you supply sufficient leaf material to allow for this.

LGC produce a ‘DNA extraction leaf tissue collection kit’ (KBS-9370-110) that includes a 6 mm leaf punch, a leaf punching mat, the required number of 96-well sample plates with perforated strip lids, and a transport box. If you prefer to use your own plates to send in leaf tissue samples for DNA extraction, this is not a problem. You will need to use polypropylene deep-well 1.2 ml plates that are compatible with all of our laboratory procedures. We recommend that you send in an additional empty plate (either in advance of your samples or alongside your sample plates) as this can be used to test that the plate type is sufficiently robust to withstand our laboratory procedures without risk to your samples.

The leaf tissue discs must be dried, ideally by lyophilisation, prior to sealing of the plate to prevent samples from deteriorating during transit. Following addition of the dried tissue samples to the plate, ensure that each well is sealed to prevent contamination of the wells and to avoid samples moving location during transit. Please do not place silica pellets directly in the wells of the plate as these will have to be manually removed prior to performing DNA extraction and will therefore incur an additional charge.

LGC are able to extract DNA from FTA cards. FTA cards are made of a filter-paper type material and have been developed for the collection and storage of DNA from organic samples such as blood and cultured cells. Researchers apply their sample to the card and chemicals within the card lyse the cells, denature any proteins and preserve the remaining DNA. The extracted DNA is preserved within the card for transport or long-term storage at room temperature. To use the DNA in the laboratory, a section (punch) of the card is washed in an elution buffer. The resultant in-solution DNA can then be used as template in genotyping reactions.

The standard extraction service can take several weeks and is dependent on the project and the scheduled workload in the lab at that time.

We have received saliva samples in a range of different vessels, and these have all worked fine with our laboratory methods. Commonly used brands are the vessels from the Orangene™ DNA self-collection kit from DNA Genotek and tubes that contain a DNA preservative, such as those from Norgen Biotek Corporation.

• Whole blood
• Buffy coats *
• Buccal swabs
• Saliva OraGene tubes
• Saliva (Other)
• Serum *
• Human tissue **
• Animal tissue punctures **
• Cells / Cell cultures (please provide number of cells in case of RNA extraction)
• Plant leaves **
• Plant seeds **
• Plant roots **
• Other plant material
• Fin clips (on ethanol)
• Faecal matter
• Urine
• Paper blood spots
• Many other sample types - please contact our technical support team

* Serum and buffy coat yields can vary greatly dependent on individual samples; we advise that a pilot study is performed for a reliable prediction about the DNA yield for your specific samples.

** Please supply in 96-well plates in a sorted manner.

To normalise DNA samples in-house, LGC bring an aliquot of the neat DNA to a desired concentration (typically 50ng / µL) in a 96-well plate. Note that the standard maximum volume per normalised sample is 1 mL. Any remaining neat DNA can be shipped back to the customer where requested..

LGC can extract DNA from samples of any volume up to 50 mL. For sample volumes greater than 50 mL, customised options are available. Please contact our customer service team for more information.

LGC offers whole genome amplification (WGA) services that can be used to increase the amount of DNA. More information about WGA can be found here.

• Concentration normalisation of DNA extracts required (normalised DNA will be prepared in 96-well plates.)
• Quantification of DNA / RNA concentration (UV measurement)
• Cherry picking of samples (Most suitable from 96-well plates, or 384-well plates in some cases. Cherry picking from 1536-well plates is not feasible. Please ask your project manager or our scientific support team for more details)
• Sample transfers
• Sample management
• Sample tracking
• Plating of samples
• Plate transfer
• Additional aliquots of DNA
• Sample storage for KASP genotyping (free of charge)
• Quality check of RNA with Agilent 2100 Bioanalyzer
• Gel electrophoresis analysis
• Genotyping QC (3 KASP assays incl. gender test)
• Whole Genome Amplification (WGA)
• Genotyping projects
• Sequencing projects

Please contact us for more information.

Yes. LGC are able to offer an accredited DNA extraction service for research applications if requested (DIN EN ISO 9001:2008).

Yes, for clinical and diagnostic applications LGC offers a fully certified DNA extraction service (DIN EN ISO 9001:2008).

We would expect to obtain around 20-30 µg of DNA per 1 mL of human whole blood from a healthy individual. The DNA yield will be dependent on several factors such as the health of the individual, the technique used to obtain the sample, and the way that the sample has been stored.

The minimum volume required to perform normalisation using robotics is 60 µL.

The total DNA yield will be dependent on several factors including the health of the individual, the technique used to obtain the sample, and that way that the sample has been stored. The values below are intended as a guideline only: