PCR Cleanup Filter Plates
Remove primers and unincorporated dNTPs in one step
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Documentation
Referencias bibliográficas
| Visión general referencias | Aplicación |
|---|---|
| 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 |
Preguntas frecuentes
| Pregunta | Respuesta |
|---|---|
| My recoveries with the MultiScreen384-PCR plate are lower than expected. How can I improve them? | Here a few things to check:
|
| Will mineral oil be removed by the MultiScreen 384-PCR plate? | No. Oil will resuspend with PCR products. All 384 thermocyclers use "Hot Bonnet" technology, however, eliminating the need for oil. |
| When using the Multiscreen PCR plates, what is the minimum and maximum DNA loading capacity per well? | The minimum DNA loading capacity is one microgram per well. There is no upper limit. |
| What is the minimum recovery volume possible when using the MultiScreen-PCR plates? | The minimum recovery volume is 50µl. Use of smaller volumes can be used but will result in a decrease in DNA recovery. |
| What are the dimensions of the Multiscreen-PCR plate? | The plate length is 123.4mm. The plate width is 82.7mm. the plate depth is 14.6mm without the cover. 16.5mm with cover. |
| 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. |
| What is the DNA binding capacity of the MultiScreen FB for PCR purification? | The DNA binding capacity is 4-5ug per well. |
| 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. |
Productos y aplicaciones relacionados
Productos relacionados por: Application Facete
| Protein Purification |
| Sample Preparation |
| Dialysis and Filtration |
Productos relacionados por: Brand Facete
| MultiScreen™ |
Categorías
| 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
)
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