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HighPrep HMW DNA Kit
The HighPrep High Molecular Weight DNA Kit is a magnetic bead-based kit for high molecular weight DNA isolation from whole blood, bone marrow, saliva, buccal cells, cultured cells, tissues, and bacteria.
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Step-by-Step Protocol for High Molecular Weight DNA Isolation Using the HighPrep HMW DNA Kit
Watch this step-by-step video to learn how to use the HighPrep HMW DNA Kit. This kit guides you through four steps: Lysis, DNA Binding, Bead Wash, and DNA Elution, ensuring high purity and yield (50-300+ kb).
HMW DNA isolation is suitable for molecular applications such as::
- Long read sequencing
- Next generation sequencing
- Microarray
- PCR amplification and genotyping
- Cloning
- Restriction enzyme digestion
- Isolation of high-quality, high molecular weight DNA 50-300+ kb
- HMW DNA is suitable for all third-generation sequencing platforms including Nanopore and PacBio SMRT sequencing
- Achieve high yields of DNA from a wide range of sample matrices
- Efficient scale-up and flexibility
- Amenable to automation and has easy to use protocols (Kingfisher™ scripts are ready)
- Comes with RNase to minimize the co-purification of RNA
The HighPrep High Molecular Weight DNA Kit is specifi cally designed for isolation of high molecular weight DNA (HMW DNA) in the size range of 50–300+ kb. The kit extracts HMW DNA from biological samples such as whole blood, bone marrow, saliva, buccal cells, cultured cells, tissues, and bacteria. The HighPrep High Molecular Weight DNA Kit utilizes magnetic bead-based technology in combination with chaotropic agents to gently isolate HMW DNA and remove inhibitors. The HMW DNA purifi ed with this kit is particularly suitable for analysis on long read sequencing genomic platforms including PacBio RSII/Sequel/Sequel II and Oxford Nanopore. DNA purifi ed using the HighPrep High Molecular Weight DNA Kit is of good quality and purity. The kit includes RNase A for the removal of RNA. The exact size of extracted DNA varies depending on sample matrix, the quality of the starting material, and processing conditions.
This is a short overview of the protocol. For a detailed protocol, please refer to PRODUCT DOCUMENTS.
- Pre-Processing (optional depending on the sample type): Homogenization of solid tissues, centrifugation or pelleting of cells from collection or growth medium.
- Lysis: Cell lysis, releasing DNA and RNA into the solution. Cell lysis is carried out by HAS Buff er, HTS Buff er and Pro K. RNase A degrades RNA in the sample
- DNA Binding: MAG-HM1 Particles in the presence of 100% Ethanol and a chaotropic salt bind the DNA in the solution.
- Bead Wash: Wash of MAG-HM1 Particles with HMW1 Wash Buffer and HMW2 Wash Buffer
- DNA Elution: Addition of a low molarity Buffer of a basic pH releases DNA from the beads into the Elution Buffer
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Starting Material (Diverse Sample Types) |
200 μL of blood, bone marrow, and saliva, up to 10 mg of tissue, up to 1 x 10^9 Cultured cells, bacterial pellet with OD600=1 or less, 200 μL buccal swab cells in medium |
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Process Method |
Supports manual and automated processing |
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Instrument |
Adaptable to most nucleic acid purification instruments, but a ready-made script for Kingfi sher™ Flex is available upon request. |
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Purification Method |
Magnetic bead-based technology |
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Elution Volume |
30-100 μL |
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Size Range |
50-300+ kb |
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Purity |
A260/A280 and A260/A230 of >1.7 |
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Yield |
Depends on the quality and amount of sample. 10 mg of tissue yields 10 μg of DNA or higher. 200 μL of human whole blood yields 3-7 μg of DNA. |
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Automated Extraction on Kingfi sher™ Flex |
1 hr. 20 mins |
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Manual Extraction |
2 hrs. |
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Downstream Applications |
Long read sequencing, Next generation sequencing, Real-time quantitative PCR, Reverse transcriptase PCR, PCR, Microarray, Genotyping, Restriction enzyme digestion |