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: | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | A | B2436 34yr | BD2445 69yr | BD2617 36yr | BD2449 38yr | BD2465 40yr | BD2547 54yr | BD2561 53yr | BD2538 40yr | BD2565 34yr | BD2458 28yr | BD2638 34yr | BD2644 39yr | B | BD2535 34yr | BD2678 50yr | BD2671 46yr | BD2494 45yr | BD2550 49yr | BD2560 37yr | BD2481 35yr | BD2415 35 | BD2416 39 +42yr | BD2439 31yr | BD2647 35yr | BD2500 46yr | C | BD2536 26yr | BD2531 52yr | BD2530 38yr | BD2491 49yr | BD2499 44yr | BD2443 36yr | BD2654 53yr | BD2599 39yr | BD2662 33yr | BD2613 33yr | BD2532 39yr | BD2632 46yr | D | BD2418 37yr | BD2685 37yr | BD2426 28yr | BD2437 42yr | BD2438 40yr | BD2590 37yr | BD2603 54yr | BD2586 29yr | BD2605 42yr | BD2612 44yr | BD2639 41yr | BD2633 50yr | E | BD2670 45yr | BD2446 34yr | BD2447 43yr | BD2464 39yr | BD2467 50yr | BD2598 38yr | BD2566 52yr | BD2572 32yr | BD2569 35yr | BD2585 39yr | BD2640 42yr | BD2634 59yr | F | BD2473 37yr | BD2549 35yr | BD2552 33yr | BD2679 39yr | BD2487 54yr | BD2579 33yr | BD2591 58yr | BD2568 40yr | BD2595 31yr | BD2596 48yr | BD2642 48yr | BD2643 27yr | G | BD2488 43yr | BD2539 50yr | BD2559 26yr | BD2525 28yr | BD2564 37yr | BD2584 41yr | BD2567 30yr | BD2663 42yr | BD2665 54yr | BD2645 36yr | BD2645 36yr | BD2646 26yr | H | BD2548 42yr | BD2540 40yr | BD2553 32yr | BD2546 32yr | BD2537 34yr | BD2614 34yr | BD2618 39yr | BD2619 28yr | BD2620 47yr | BD2628 55yr | BD2655 38yr | BD2648 38yr |
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.
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