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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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