Reagents & Equipment (go back)
This is intended to provide a complete listing of reagents and equipment (along with specific vendors and catalog numbers) for everything required to get going with the polony technology. Again, we strongly urge users to stick to this reagent and equipment list, ordering the specific items listed from the specific vendors listed, and to only make substitutions after having clearly established a working implementation of the protocol. This will improve the likelihood that all will work as predicted the first time around, and aids in troubleshooting if it does not. There are of course obvious exceptions to this (e.g. the brand of your flat shaker). The items listed here (along with the Starters Kit, if you choose to use it), should be all that you need to get started. Costs are approximate. If you notice reagents or equipment named in publications or experimental protocols that are not mentioned here, please let us know.
A microarray scanner is far and above the most expensive item on this list. To date, we only have experience with Perkin-Elmer and Axon instruments. We encourage you to take advantage of whatever equipment you have access to, and would be happy to send you test-slides (e.g. gels that are already amplified and labeled) if you wish to test the suitability of your scanning equipment.
Contact us if you are interested in the nucleotides with cleavable labels (e.g. Cy5-SS-dNTPs or Cy5-PC-dNTPs).
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GEL COMPONENTS
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| 40% IEF Acrylamide (1000 mL) | Amersham Biosciences | 17-1301-01 | 86.00 | |
| Acrylamide / Bis (19:1; 38%:2%) | Roche | 1-814-290 | 77.00 | |
| DATD ((+)-N,Ną-diallyltartramide), 25 g | Sigma-Aldrich | 15,686-8 | 31.50 | |
| APS (100 g) | Sigma-Aldrich | A-7460 | 35.10 | |
| TEMED (25 mL) | Sigma-Aldrich | T-9281 | 18.00 | |
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SLIDES & SLIDE HANDLING
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| Teflon Coated Glass Slides (100 / pack) | Erie Scientific | 285W [white coat] | 35.00 | 4 to 6 weeks for delivery is typical. |
| Glass Cover Slips (18 mm x 30 mm, #1) | Fisher Scientific | CUSTOM | 300.00 | 4 to 6 weeks for delivery is typical. |
| 125 uL FrameSeal Chambers (100 / pack) | MJ Research | SLF-1201 | 78.00 | 4 to 6 weeks for delivery is typical. |
| SA500 SecureSeal Chambers (50 / pack) | Grace Biolabs | 10484915 | 142.00 | 4 to 6 weeks for delivery is typical. |
| Plastic Coplin Jars (each) | VWR | 25457-200 | 5.75 | |
| Glass Coplin Jars (6) | Sigma-Aldrich | S-5516 | 119.90 | |
| Black Box Slide Box | Research Products Intl. | F-44182 | 33.50 | |
| Black Box Slide Rack | Research Products Intl. | F-44180 | 11.75 | |
| Molded Plastic Slide Storage Box (10) | VWR | 48444-004 | 76.80 | |
| Large Glass Slide Bucket (3) | Fisher Scientific | 08-813C | 249.39 | Contact us if you find a vendor who sells these individually. |
| Metal Slide Holding Rack (6) | Fisher Scientific | 08-813A | 619.42 | Contact us if you find a vendor who sells these individually. |
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PCR & SBE REAGENTS
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| Jumpstart TAQ with 10x Buffer (w/ MgCl2) (250 U) | Sigma-Aldrich | D-9307 | 168.90 | |
| 30% BSA, 10 mL | Sigma-Aldrich | A-7284 | 23.85 | |
| Klenow Polymerase (3'-5' exo-minus) | NEB | M0212M | 200.00 | High Concentration (50K Units / mL) |
| SSB (100 ug) | USB | 70032Y | 84.00 | |
| 10 mM unlabeled dNTP mix (100 uL) | Gibco-BRL | 18427-013 | 54.00 | |
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SOLUTIONS
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| Hexane (500 mL) | Sigma-Aldrich | H-9379 | 39.90 | |
| Tris-Hydrochloride | JT Baker | 4103-04 | 49.50 | |
| Trizma Base (500 g) | Sigma-Aldrich | T-6066 | 46.65 | |
| Acetic Acid, 99.5% Glacial (500 mL) | Mallinckrodt | UN2789 | 17.30 | |
| Formamide (250 mL) | Sigma-Aldrich | F-5786 | 50.40 | |
| Iodoacetamide (5 g) | Sigma-Aldrich | I-1149 | 27.40 | |
| 2-mercatpoethanol (100 mL) | Sigma-Aldrich | M-6250 | 14.70 | |
| Tween-20 (100 mL) | Sigma-Aldrich | P-1379 | 12.30 | |
| Triton X-100 (50 mL) | Sigma-Aldrich | T-8787 | 13.10 | |
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LABELED NUCLEOTIDES
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| Cy5-dATP | Perkin Elmer / NEN | Contact Vendor | ??? | Renaissance Product Line |
| Cy5-dGTP | Perkin Elmer / NEN | Contact Vendor | ??? | Renaissance Product Line |
| Cy5-dCTP (25 nmols) | Amersham Biosciences | PA-55021 | 295.00 | 100 nmols can be had for $475.00 |
| Cy5-dUTP (25 nmols) | Amersham Biosciences | PA-55022 | 295.00 | 100 nmols can be had for $475.00 |
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OTHER
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| Argon Gas | MedTech | 11J12 | 30.00 | |
| Bind Silane (25 mL) | Amersham Biosciences | 17-1330-01 | 80.00 | |
| Sybr Green I (500 uL, 10,000x) | Molecular Probes | S-7563 | 149.00 | |
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MISCELLANEOUS
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| Sharpie Pen | Staples | 495190 | 1.59 | |
| 0.22 micron filters (150 / pack) | Sigma-Aldrich | Z35,549-6 | 239.80 | |
| 3cc Syringes (100 / pack) | Becton-Dickinson | 309585 | 15.10 | |
| Tweezers (several pairs) | ||||
| Razor Blades | ||||
| 15 mL polypropylene conicals | ||||
| ddH20 (autoclaved) | ||||
| Stir Bars | ||||
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EQUIPMENT
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| AirClean 600 PCR Workstation | AirClean Systems |
??? |
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| ScanArray Express HT with Autoloader | Perkin Elmer Life Sciences | 80,000.00 | ||
| Thermal Cycler | MJ Research | ALD-0211 | ??? | |
| Slide Cycling "16/16" Dual Block | MJ Research | PTC-0200 | ??? | |
| Desiccation Chamber | ||||
| Flat Shakers (one at RT and one at 70'C) | ||||
| 70'C Incubator |