Human Genetic Disease DNA Panel Breast Cancer-1 Product Data Sheet	Manufactured by:
Product No: HGD BC-1 (96 Array)	Distributed by:

Intended Use
ECACC Human Genetic Disease (HGD) DNA Panels are intended for use in human genetic research. They are not intended as a primary source of case control DNA but to provide a readily available source of DNA which may serve as a verification panel to use against your own case control set. The DNA arrays are designed for single use and can be used directly in automated gene analysis systems.



Description
HGD BC-1 DNA is extracted from lymphoblastoid cell lines derived by Epstein Barr Virus (EBV) transformation of peripheral blood lymphocytes from fresh, patient blood samples deposited by a single centre, Guys Hospital, London. The cases are female patients that have all been diagnosed as having breast cancer. Where the age of first diagnosis was known it is indicated in the table below. The type of mutation affecting these patients is not known. All donors have given written, informed consent for their blood to be used for research purposes.

The DNA is extracted and purified using standard techniques that yield high molecular weight material, as verified by agarose gel electrophoresis. The composition of each array is completely defined and standardised so that each product lot will be identical. The identity of each product lot is checked using STR DNA profiling. Consequently ECACC HGD DNA Panels can be used as Reference Standard material as an aid to routine quality control in the laboratory.


Sample details
The table below provides the individual sample designation and age of patient at diagnosis:

Amount and concentration of DNA
The DNA concentration is measured using the ds DNA Quantitation kit (Molecular Probes, USA) on a Fluroskan Ascent from Labsystems (Type 374).

The DNA is dissolved in 10mM Tris (pH8) 1mM EDTA to preserve the quality of the DNA.

Total DNA per sample:  Minimum of 400ng
Concentration:              Target 8ng/µl Typical range +/- 2ng/µl
Recoverable volume:    50µl


Storage
ECACC DNA array are supplied frozen to preserve the quality and also to facilitate handling. On receipt, panels should be stored at -80°C. The DNA array presentation is intended for single use although, with care, repeat use is possible. After thawing, panel can be stored at 2-8 °C for up to 6 months, provided precautions are taken to prevent fluid loss by evaporation.

Repeated freeze-thawing cycles can damage the DNA and should be avoided.


Use of DNA Arrays
Remove the array from its polythene bag / packaging and make sure this Data Sheet is retained safely. Do not remove the foil seal.

Place the array on a level surface, top uppermost and allow to thaw. Centrifuge the plate to force all the fluid to the bottom of the well. Do not remove the foil seal until these precautions have been observed.

Important - If it is not possible to centrifuge the plates then the foil seal must be removed immediately after removal from freezer storage. (If the plate is allowed to thaw, liquid may condense on the underside of the foil seal.)

Alternatively the foil seal may be pierced using a sharp point to allow access to micropipette tips. This approach minimises the risk of cross contamination associated with the action of peeling off the foil seal.

Use a micropipette to withdraw the required amount of DNA solution from the array.

Although the arrays are designed for single use they can be resealed using the replacement foil seal included and returned to store. (See advice on storage, above).


Warnings and Precautions
Avoid cross contamination between different samples comprising an array. Do not repeatedly use the same foil seal. Remove adhesive sealers with care. Use clean pipette tips for each specimen.

The master cell banks used to prepare the DNA extracts are taken from 'low risk' normal blood donors. However it is recommended that standard laboratory precautions for handling potentially infectious material should be employed.


Limitations of Use
ECACC DNA Panels are intended for research use only. The diagnostic data provided is that which ECACC received with the original blood sample from the original depositor and has not been confirmed by ECACC.