PCR Cleanup Filter Plates
Remove primers and unincorporated dNTPs in one step
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Panoramica
Specifiche tecniche
Informazioni per l’ordine
Documentation
Riferimenti bibliografici
| Panoramica dei riferimenti bibliografici | Applicazione |
|---|---|
| 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 1999 | |
| 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) 1999 | 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 1994 | Nucleic Acid Purification and Concentration |
FAQ
| Domanda | Risposta |
|---|---|
| When using the Montage kits how can I tell when filtration in the MultiScreen plate is complete? | Filtration is complete when the individual wells of the MultiScreen plate are empty and the membranes appears shiny. Filtration usually takes about 10 minutes, depending on starting volume. |
| When using the Montage PCR-96 kit, what is the recommended procedure for resuspension of PCR fragments post filtration? | Avoid manual pipetting. The use of a plate shaker or automated pipettor (on robotic liquid handling system) is highly recommended. |
| What is the minimum resuspension volume for product recovery from the Montage PCR96 Kit? | 50 microliters per sample. |
| What is the maximum binding capacity for DNA in the PCR Product Purification protocol using the MultiScreen-FB plate? | The binding capacity of the MultiScreen FB, after treatment with "binding" buffer is 4-5ug of DNA per well. |
| Can the Montage PCRm96 plate be used in centrifugal mode? | No. The Montage PCRm96 plate is designed for purification using vacuum filtration only. |
| Will the Montage PCR96 and Montage PCRm96 plates remove primers and primer-dimers? | The Montage PCR96 Cleanup Kit is designed to remove primers and other low molecular weight material from the PCR reaction. However, removal of primer dimers is problematic. The Montage PCRm96 plate removes > 99% of PCR primers, but although small primer-dimers are efficiently removed, quantitative removal of larger primer-dimers can be problematic given that the PCRm96 plate has been optimized for recovery of small PCR products. The best way to avoid complications due to primer-dimers is to optimize primer design. |
| When using Montage PCRm96 plates, do I need to do anything special to increase the recovery of my 100 - 300 bp PCR fragments? | The protocol that was developed in conjunction with the PCRm96 plates has been optimized to provide maximal recovery of even small PCR products (e.g., 100 – 300 bp). No special modifications are necessary for small PCR fragments. |
| If I dilute my sample when using the Montage PCRu96 plate, will that increase the amount of time for filtration to be completed? | Yes. Although sample dilution will marginally increase processing time, dilution is essential for maximal recovery of PCR products. |
| Is a wash step required following the initial filtration when using the Montage PCRu96 plate? | The PCR product is sufficiently pure for DNA sequencing after a single vacuum filtration. If a particular application demands even higher purity, a wash step may be added. |
| What is the maximum vacuum that can be applied to the Montage PCRu96 plate? | The recommended vacuum pressure for PCR purification is 20 inches of Hg. Exceeding this vacuum pressure will adversely affect recovery of PCR products and is not recommended. |
Prodotti e applicazioni correlate
Prodotti correlati per: Application Facete
| Protein Purification |
| Sample Preparation |
| Dialysis and Filtration |
Prodotti correlati per: Brand Facete
| MultiScreen™ |
Categorie
| 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
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| 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 |
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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
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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