Isolate genomic DNA from animal tissues, cells, bodily fluids, viruses and swabs
Rapid and convenient spin column procedure
Purified DNA is of the highest quality and integrity for sensitive downstream applications including PCR, qPCR, genotyping, sequencing and more
This kit is designed for the rapid preparation of genomic DNA from various tissue samples, cultured cells, viruses, bodily fluids and swabs using a rapid spin column protocol. Purified DNA is of an excellent yield and quality, and is immediately ready for any downstream application including PCR, qPCR, genotyping, sequencing and more. The protocol can be completed in approximately 80 minutes (including incubation time).
Average Yield:* HeLa Cells (1 x 106) Tissue (from 10 mg kidney)
8 µg 10 µg
Maximum Amount of Starting Material: Animal Tissues Cultured Cells Bodily Fluids (blood, saliva) Viral Suspension
20 mg 3 x 106 cells 150 µL 150 µL
Time to Complete 10 Purifications
80 minutes
* Yield will vary depending on the type of sample processed
Storage Conditions and Product Stability The Proteinase K should be stored at -20°C upon arrival and after reconstitution. All other solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment.
These kits provide a fast, reliable and convenient spin column method for the isolation of high quality, high purity and inhibitor-free cell-free circulating DNA (cfc-DNA) from plasma/serum sample. These kits are designed to isolate all sizes of cfc-DNA from either fresh or frozen plasma/serum samples and the purified DNA is eluted into a flexible elution volume ranging from 25 µL to 50 µL. The purified plasma/serum cfc-DNA is fully compatible with all downstream applications including PCR, qPCR, methylation-sensitive PCR and Southern Blot analysis, microarrays and NGS.
Background
Plasma/Serum cell-free circulating DNA (cfc-DNA) has the potential to provide biomarkers for certain cancers and disease states as well as fetal DNA in maternal blood. Currently, significant advancements are being made in utilizing cfc-DNA as biomarkers for the early diagnosis, prognosis and monitoring of therapy for several cancer types and autoimmune diseases. Cell-free mitocondrial DNA (cfmtDNA) is also under investigation for its clinical significance. This cfc-DNA is usually present as short fragments of less than 1000 bp. In addition, cell-free fetal DNA has been widely used as a non-invasive method for prenatal diagnosis including early identification of fetal sex, genetic studies for families at high risk for inherited genetic disorders, screening for Rhesus factor, screening for aneuploidy and identification of preeclampsia.
Plasma/Serum Cell-Free Circulating DNA Purification Micro Kit Dx
For serum input volumes 10 µL – 200 µL. Purify high-quality DNA in 15-20 minutes
Plasma/Serum Cell-Free Circulating DNA Purification Mini Kit Dx
For serum input volumes 200 µL – 500 µL. Purify high-quality DNA in 15-20 minutes.
Plasma/Serum Cell-Free Circulating DNA Purification Midi Kit Dx
The first column will handle the large volume input of bodily fluids that is followed by a concentration on a mini column for a final elution of 25 µL to 50 µL. All components for the purification & concentration are provided in one convenient & fast kit for the easy processing of large input volumes of bodily fluids. For a schematic workflow of the protocol click here to view. For serum input volumes 1 mL – 4 mL. Purify high-quality DNA in 90 minutes.
Plasma/Serum Cell-Free Circulating DNA Purification Maxi Kit Dx
The first column will handle the large volume input of bodily fluids that is followed by a concentration on a mini column for a final elution of 25 µL to 50 µL. All components for the purification & concentration are provided in one convenient & fast kit for the easy processing of large input volumes of bodily fluids. For a schematic workflow of the protocol click here to view. For serum input volumes 5 mL – 10 mL. Two kits in one – purify and concentrate DNA from bodily fluids using a convenient two column system.
*Please check page 7 in the product manual for the Plasma/Serum Average Yields and the Common DNA Quantification Methods.
Storage Conditions and Product Stability All buffers should be kept tightly sealed and stored at room temperature (15-25°C) for up to 2 years without showing any reduction in performance. Norgen’s Plasma/Serum Cell-Free Circulating DNA Purification Kits contain ready-to-use Proteinase K solution, which is dissolved in a specially prepared storage buffer. The Proteinase K is stable for up to 2.5 years after delivery when stored at room temperature. To prolong the lifetime of Proteinase K, storage at 2–8°C is recommended.
The HiPure Fastfilter Plasmid DNA Midi Kits combine the power of HiPure technology with the time-tested consistency of alkaline-SDS lysis of bacterial cells to deliver high-quality DNA. HiPure DNA columns facilitate the binding, washing and elution steps thus enabling multiple samples to be simultaneously processed. This system also includes aspecial filter cartridge which replaces the centrifugation step following alkaline lysis. Plasmid DNA purified by this system is suitable for automated fluorescent DNA sequencing, restriction endonuclease digestion, transfection of mammalian cells, and other manipulations. Up to 250 µg high copy number plasmid DNA or 10-75 µg low copy number plasmid DNA can be purified from 15-50 mL overnight culture.
Details
Specifications
Features
Specifications
Main Functions
Isolation up to 250μg plasmid DNA from 30-50ml bacterial culture
Applications
Enzyme digestion, sequencing, PCR and labeling, etc.
Purification method
Midi spin column
Purification technology
Silica technology
Process method
Manual (centrifugation or vacuum)
Sample type
Conventional plasmid, plasmid less than 30KB
Sample amount
30-50ml
Yield
50-250µg
Elution volume
≥300μl
Time per run
≤60 minutes
Liquid carrying volume per column
4ml
Binding yield of column
250µg
Principle
The HiPure Plasmid procedure is based on alkaline lysis of bacterial cells followed by adsorption of DNA onto silica in the presence of high salt. The unique silica membrane used in the kit completely replaces glass or silica slurries for plasmid DNA minipreps. The procedure consists of 3 basic steps: Preparationand clearing of a bacterial lysate by alkaline method，then transfer the supernatant to column to bind DNA. After washing proteins and other impurities, nucleic acid was finally eluted with low-salt buffer (10mm Tris, pH9.0, 0.5mm EDTA).
Advantages
High purity – purified plasmid can be directly used in sequencing, enzyme digestion and PCR, etc.
Fast – it takes only 15-60 minutes to complete the isolation
High yield – up to 70µg plasmid can be binded in one column
Kit Contents
Contents
P101302
P101303
Purification Times
25 Preps
100 Preps
RNase A
10 mg
40 mg
Buffer P1
80 ml
300 ml
Buffer P2
80 ml
300 ml
Buffer LEN3
40 ml
150 ml
Buffer GBT
60 ml
250 ml
Buffer PW1
60 ml
250 ml
Buffer PW2
20 ml
100 ml
Elution Buffer
30 ml
120 ml
HiPure DNA Midi Columns III
25
100
Lysate Clear Midi Syringe
25
100
15 ml Collection Tubes
50
200
Storage and Stability
The kit components can be stored dry at room temperature (15-25°C) and are stable for at least 18 months under these conditions. If any precipitates form in the buffers, warm at 37℃ to dissolve. After addition of RNase A，Buffer P1 is stable for 6 months when stored at 2-8°C.
The HiPure Fastfilter Plasmid DNA Midi Kits combine the power of HiPure technology with the time-tested consistency of alkaline-SDS lysis of bacterial cells to deliver high-quality DNA. HiPure DNA columns facilitate the binding, washing and elution steps thus enabling multiple samples to be simultaneously processed. This system also includes aspecial filter cartridge which replaces the centrifugation step following alkaline lysis. Plasmid DNA purified by this system is suitable for automated fluorescent DNA sequencing, restriction endonuclease digestion, transfection of mammalian cells, and other manipulations. Up to 250 µg high copy number plasmid DNA or 10-75 µg low copy number plasmid DNA can be purified from 15-50 mL overnight culture.
