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.

APPLICATIONS

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

BENEFITS

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

PROTOCOLS

This is a short overview of the protocol. For a detailed protocol, please refer to PRODUCT DOCUMENTS.

1. Pre-Processing (optional depending on the sample type): Homogenization of solid tissues, centrifugation or pelleting of cells from collection or growth medium.
2. 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
3. DNA Binding: MAG-HM1 Particles in the presence of 100% Ethanol and a chaotropic salt bind the DNA in the solution.
4. Bead Wash: Wash of MAG-HM1 Particles with HMW1 Wash Buffer and HMW2 Wash Buffer
5. DNA Elution: Addition of a low molarity Buffer of a basic pH releases DNA from the beads into the Elution Buffer

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
Process Method	Supports manual and automated processing
Instrument	Adaptable to most nucleic acid purification instruments, but a ready-made script for Kingfi sher™ Flex is available upon request.
Purification Method	Magnetic bead-based technology
Elution Volume	30-100 μL
Size Range	50-300+ kb
Purity	A260/A280 and A260/A230 of >1.7
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.
Automated Extraction on Kingfi sher™ Flex	1 hr. 20 mins
Manual Extraction	2 hrs.
Downstream Applications	Long read sequencing, Next generation sequencing, Real-time quantitative PCR, Reverse transcriptase PCR, PCR, Microarray, Genotyping, Restriction enzyme digestion

LEARN ABOUT OTHER PRODUCTS