ECACC Nucleic Acids

Section Overview

• Cell Lines: Important Tools for Elucidating the Role of Genes in Disease and Therapeutics
• Ready-to-assay Nucleic Acids from Cell Lines for Appropriate Model Choice
• Genomic DNA from Cell Lines
• Quality Control for Cell Line DNA
• Human Random Control DNA Panels
• How to Handle Cell Line DNA on Arrival
• Advantages of Obtaining Corresponding Nucleic Acids from ECACC

Cell lines: important tools for elucidating the role of genes in disease and therapeutics

Genomic DNA is available as a standard items on each cell line product page for cell lines where it is available. We additionally provide a range of Human Random Control DNA panels. These comprise 96 control DNA samples (2 mg of DNA at a concentration of 100 ng/mL) supplied in a 96-well plate. Five different panels are available.

Genomic DNA from cell lines

We offer high quality, high molecular weight genomic DNA extracted from ECACC cell lines. To ensure the highest quality and reproducibility, extracted DNA is subjected to defined quality control procedures, and all ECACC DNA products are supplied at a standard concentration of 100 ng/μL in 5 µg per tube.

Genomic DNA is extracted from cell lines using an automated extraction system employing silica-coated magnetic bead technology. All DNA is quantified using the A260 absorbance assay and normalized to a standard concentration of 100 ng/µL.

Quality control for cell line DNA

To ensure the highest quality, all ECACC DNA products are subjected to defined quality control procedures which include assays for:

• DNA Size and Integrity: Agarose gel electrophoresis is used to verify the molecular weight and integrity of cell line DNA. Our extraction procedure consistently produces genomic DNA between 20-30 kb in size.
• Purity: 260/280 nm and 230/260 nm UV spectrophotometry ratios are used to determine purity of the extracted DNA
• Authenticity: Short tandem repeat (STR) PCR, using a commercially available forensic kit consisting of 10 markers plus amelogenin (a marker for gender determination), is used to verify the identity of each batch of DNA against source material.
• Quantification: PicoGreen® fluorescence assay is used to quantify DNA
• Performance: DNA is independently assessed using an assay selection that includes HLA typing, STR PCR, 5’ nuclease assay, and more
• Long term stability and performance: Long term stability and performance assays of DNA samples from the Human Random Control DNA panels involved repeated agarose gel electrophoresis as well as the repeated use in PCR analysis over several years. There was no reduction of PCR product observed for samples that were properly stored for up to 28 months at 4˚C.

Human Random Control DNA Panels

Human Random Control (HRC) DNA is a readily available, cost-effective, and renewable source of authenticated, high quality purified genomic DNA from a control population of 480 randomly selected, non-related UK Caucasian blood donors. DNA is extracted from lymphoblastoid cell lines from single donor blood samples derived by EBV (Epstein-Barr virus) transformation of peripheral blood lymphocytes and is available as a series of five panels. Each panel contains samples from 96 unique individuals in a convenient 96-well format. DNA is provided at a concentration of 100 ng/μL in 10 mM Tris buffer (pH 8.0) with 1mM EDTA. Gender and age at blood collection is available for most samples.

How to handle cell line DNA on arrival

Frozen DNA products should be stored at -20 °C upon receipt if they are not used immediately. Repeated freeze/thawing should be avoided. On receipt of your DNA order, ECACC recommends the following:

DNA Supplied in Tubes

1. Once thawed, the DNA can be stored at 4 °C to 8 °C for at least six months
2. Before removing the cap, place the tube in a mini-centrifuge for a short spin to bring contents to the bottom of the tube
3. To prevent cross-contamination, take care not to mix caps between samples. If in doubt, use a fresh cap.

DNA Supplied in 96-well Plates

1. Once the plates have been thawed, and prior to piercing the seal, they should be centrifuged briefly to bring all contents to the bottom of the plate wells
2. DNA supplied in 96-well plates is intended for single use. However, should it be necessary to store the plates, the plastic lid should be placed over the pierced wells. The plate should be stored upright at 4 °C to 8 °C. The DNA will evaporate rapidly if the plate is not re-sealed using the included replacement seal. When resealed, the DNA can be stored for at least three months at 4 °C to 8 °C.
3. ECACC does not recommend refreezing the plates.

Advantages of obtaining corresponding nucleic acids from ECACC

For cell lines where genomic DNA is available, it is available on the corresponding cell line product page.