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HighPrep PCR PB
Note: This product is not available for purchase in Europe
HighPrep PCR PB is a paramagnetic bead-based product, specifically designed for SMRT sequencing workflow. It is used for DNA cleanup and size selection of SMRTbell® libraries.
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Step-by-Step Protocol for DNA Purification and Size Selection Using HighPrep PCR PB
Watch this step-by-step video to learn how to use the HighPrep PCR PB. This enhanced replacement of AMPure PB isolates and purifies high-quality DNA, ensuring it is free from contaminants, and precisely selecting the desired DNA fragment sizes for your applications. Follow along to achieve optimal results for HiFi PacBio Long-Read Sequencing.
HighPrep PCR PB is suitable for PacBio applications such as:
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SMRTbell® libraries clean-up and size selection
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Enhanced direct replacement of AMPure PB, no protocol
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Cost effective. Save at least 40% or more when compared with the competitor PB
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Amenable to automation (Kingfisher™ Flex script ready)
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Purifies starting DNA material with low purity (low A260/A230) to remove contaminants
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Efficiently removes unincorporated dNTPS, primer dimers, excess primers, salts, and other impurities from amplicons
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Concentration of DNA from highly diluted samples
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Depletes DNA fragmentsless than 5 kb
HighPrep PCR PB is a paramagnetic bead-based kit specifically designed for SMRT sequencing workflow. It is used for DNA cleanup and size selection of SMRTbell® libraries.
HighPrep PCR PB is free of reagents that can interfere with polymerase activity during sequencing, therefore, templates bind properly to the polymerase. This product is designed for both manual and fully automated purification of DNA and sequencing products.
This is a short overview of the protocol. For a detailed protocol, please refer to PRODUCT DOCUMENTS.
DNA Clean-up/Purification
- Add 0.45x HighPrep PCR PB reagent to the sample
- Bind DNA to paramagnetic beads
- Separation of beads from the supernatant with contaminants
- Wash beads with Ethanol to remove contaminants
- Elute purified DNA from beads
Fragment Selection
- Dilute HighPrep PCR PB to 35% v/v
- Add HighPrep PCR PB reagent (based on desired fragment size) to the sample
- Bind DNA to paramagnetic beads
- Separation of beads from the supernatant with contaminants
- Wash beads with Ethanol to remove contaminants
- Elute purified DNA from beads
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Nucleic Acid Input |
Fragmented DNA, NGS and TGS libraries, PCR products |
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Starting Amount |
Scalable |
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Recovery |
>80% |
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Process Method |
Automated; Manual |
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Instrument |
Adaptable to most nucleic acid purification instruments, but a ready-made script for Kingfisher™ Flex is available upon request |
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Elution Volume |
10 uL or above |
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Cleanup |
Removes unwanted reaction components to purify DNA |
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Size Selection |
Adjustable bead ratio based on desired fragment size |
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Automated Extraction on Kingfisher Flex |
45 minutes |
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Downstream Applications |
Genomic DNA cleanup, NGS library cleanup, Size selection, PCR, SMRTbell® libraries clean-up and size selection |