PCR Cleanup Filter Plates
Remove primers and unincorporated dNTPs in one step
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Overview
Specifications
Ordering Information
Documentation
References
| Reference overview | Application |
|---|---|
| PCR Amplification on a Microarray of Gel-Immobilized Oligonucleotides: Detection of Bacterial Toxin- and Drug-Resistant Genes and Their Mutations Strizhkov, B.N. Drobyshev, A.L. Mikhailovich, V.M. Mirzabekov, A.D. BioTechniques 29:844-857 2000 | |
| A Concise Guide to cDNA Microarray Analysis P. Hegde, R. Qi, K. Abernathy, C. Gay, S. Dharap, R. Gaspard, J.E. Hughes, E. Snesrud, N. Lee, and J. Quackenbush BioTechniques Vol. 29, No. 3: pp 548-562 (Sep 2000) 2000 | PCR |
| A specific and sensitive PCR assay suitable for large-scale detection of toxigenic Pasteurella multocida in nasal and tonsillar swabs specimens of pigs Kamp, E.M. etal., J.Vet Diagn Invest 8:, 304-309 J.Vet Diagn Invest 8:, 304-309 1996 | Nucleic Acid Purification and Concentration |
| Use of MultiScreen Plates for the Preparation of Bacterial DNA Suitable for PCR Reek, F.H., Smits, M.A., Kamp, E. M., Smith, H.E. BioTechniques. 19: 2, 282 – 285 1995 | Nucleic Acid Purification and Concentration |
FAQ
| Question | Answer |
|---|---|
| Can the Montage PCR-96 Kit be used in a procedure employing centrifugal rather than vacuum filtration? | Use of the Montage PCR-96 Kit in a centrifugal protocol is not recommended. |
| Can the MultiScreen plates included with the Montage kit be used with a vacuum manifold from a vendor other than Millipore? | Yes, but only if proper underdrain support grid is employed. Please contact your local Millipore technical service organization for specific information on the availability of the "underdrain supports" for non-Millipore vacuum manifolds. |
| Is a wash step required following the initial filtration with the Montage PCR-96 Cleanup Kit? | For many applications, a wash step is not necessary. If you need wash the purified PCR products, use the nuclease free water included in the PCR cleanup kit. |
| Should I use water or Tris buffer to resuspend the final PCR products? | Either solution may be used with the Montage PCR-96 Kit. A user may prefer one of these depending on the downstream application. |
| What are the minimum and maximum loading capacities for the MultiScreen plates used with the Montage PCR-96 Kit? | There is a 1 ug minimum capacity and no maximum limit. |
| What is filtration time for the the Montage PCR-96 Kit? | Filtration time is approximately 10 min, depending on volume of sample. |
| Is the Montage PCR-96 kit capable of eliminating Triton or other detergents from the buffers used in PCR reactions? | It is difficult to completely remove Triton. We recommend the use of a PCR buffer that is Triton-free. Alternatively, one can use MultiScreen FB plates with TN063 protocol. |
| Can the MultiScreen-PCR plate be used in a centrifugal mode? | The MultiScreen-PCR plates are designed for use in the vacuum mode and use in the centrifugal mode is not recommended. |
| Can the MultiScreen-PCR plate be used with another manufacturers vacuum manifold? | No, the plate needs the support grid of the Millipore vacuum manifold. |
| How can I increase the recovery of my 100-200 BP PCR fragment in the PCR "clean-up" application? | Millipore recommends use of the MultiScreen FB plate for "clean-up" of PCR fragnents of this size, especially with low concentrations of DNA fragments. |
Related Products & Applications
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Related Products By: Brand Facete
| MultiScreen™ |
Categories
| Life Science Research > Cell Culture and Systems > Cell Culture Flasks, Plates, & Slides > Multiwell Plates |
Our PCR filter plates are fast, automatable solutions for high-throughput PCR purification. Available in 96- and 384-well formats and a µ96-well format for small volumes, MultiScreen® PCR plates use a size-exclusion membrane and vacuum filtration to provide a one-step protocol with excellent results. No centrifugation or precipitation is required. Full sample recovery from the top of the plate enhances compatibility with liquid handling systems. MultiScreen® PCRµ96 and MultiScreen®PCR384 filter plates provide high recovery on even the smallest miniaturized reactions. Working volumes <150 µL conserve reagents and save money. For volumes >150 µL, use MultiScreen®PCR96 96-well filter plate.
Features & Benefits
- High purity and high recovery
- No centrifugation or precipitation
- Fast processing times, automation compatible
- MultiScreen® PCRµ96 filter plate recommended for small fragments (1-150 bp)
Protocol
)
| 1. | Load PCR reactions. |
| 2. | Filter with Millipore vacuum manifold for 5–10 minutes or until wells are dry. |
| 3. | Add water or buffer to each well. Agitate by shaking or pipetting. Retrieve purified samples by aspiration. |
Selection Guidelines
| Specifications | |||||
| Product name | Reaction volume |
Number of samples |
Mode of operation |
Recommended applications |
Features |
| MultiScreen® PCRµ96 Filter Plate | 20 – 150 µL | 96 | Vacuum | Sequencing Genotyping Microarray |
• Highly concentrated final sample • Reaction miniaturization |
| MultiScreen® PCR96 Filter Plate | 150–300 µL | 96 | Vacuum | Microarray | • Volumetric capacity |
| MultiScreen® PCR384 Filter Plate | 20–100 µL | 384 | Vacuum | Sequencing Genotyping Microarray |
• Highly concentrated final sample • Reaction miniaturization • Ultra-high throughput |
Recovery
| Product name | Recovery of 137 bp PCR fragment* | Recovery of 301 bp PCR fragment* | Recovery of 657 bp PCR fragment* | Recovery of 1159 bp PCR fragment* | % Primer removal (20 bases), 100 µL volume |
| MultiScreen® PCRµ96 Filter Plate | + | ++ | ++ | ++ | 99.8% |
| MultiScreen® PCR96 Filter Plate | -- | + | ++ | ++ | 98.7% |
| MultiScreen® PCR384 Filter Plate | + | ++ | ++ | ++ | 99.5% |
| -- Not Recommended | |||||
| + Good Recovery | |||||
| ++ Best Recovery | |||||
| *Results will vary based on starting concentration, load, and buffers used. Data obtained using the following concentrations: 137 bp: 10 ng/µL 301 bp: 30 ng/µL 657 bp: 55 ng/µL 1159 bp: 71 ng/µL |
|||||
Performance
Get high reproducibility with MultiScreen PCR96 Filter Plate
)
Agarose gel showing reproducibility of recovery of a 500 bp PCR product with MultiScreen PCR96 filter plate.
Long Sequencing Reads
)
Representative electropherogram from a 301 bp PCR product purified using MultiScreen PCR96 filter plate (Thermo Sequenase® II dye terminator).
Suitable for Microarrays
)
Enlarged section of a 19,299 element microarray (image reproduced with permission from Dr. John Quakenbush, The Institute for Genomic Research). High quality microarrays can be manufactured using PCR products purified with MultiScreen PCR products. Reference: Hegde, P., R. Qi, K. Abernathy, C. Gay, S. Dhaarp, R. Gaspard, J.E. Hughes, E. Snesrud, N. Lee and J. Quakenbush. 2000. A concise guide to cDNA microarray analysis. BioTechniques 29: 548–562.
Applications
PCR Cleanup, Sequencing, Genotyping, Microarray, RNAi Purification, Microarray Purification, Genomic DNA Concentration