The Kit provides a high throughput of high-quality RNA from 96 samples of cells, tissues, and yeast using silica-membrane spin column plate with a binding capacity of 100ug RNA. RNA purified using the HiPure Total RNA System is ready for applications such as RT-PCR, Northern blotting, poly A+ RNA (mRNA) purification, nuclease protection, and in vitro translation.
Detail
Introduction
The Kit provides a high throughput of high-quality RNA from 96 samples of cells, tissues, and yeast using silica-membrane spin column plate with a binding capacity of 100ug RNA. RNA purified using the HiPure Total RNA System is ready for applications such as RT-PCR, Northern blotting, poly A+ RNA (mRNA) purification, nuclease protection, and in vitro translation.
Details
Principle
HiPure RNA technology simplifies total RNA isolation. Samples are first lysed and then homogenized. Ethanol is added to the lysate to provide ideal binding conditions. The lysate is then loaded onto the HiPure silica membrane and RNA binds to the silica membrane, and all contaminants are efficiently washed away. For certain RNA applications that are sensitive to very small amounts of DNA, the residual amounts of DNA remaining can be removed using a convenient on-column DNase treatment. Pure, concentrated RNA is eluted in water.
Kit Contents
Contents
R401601
R401602
Purification Times
1 x 96
4 x 96
HiPure RNA Plate
1
4
1.5ml Collection Plate
1
4
0.5ml Collection Plate
1
4
RTL Lysis Buffer
100 ml
400 ml
RNA Binding Buffer*
30 ml
120 ml
Buffer RW1
100 ml
400 ml
Buffer RW2*
50 ml
2 x 100 ml
RNase Free Water
20 ml
80 ml
Storage and Stability
The Kit can be stored dry at room temperature (15-25°C) and are stable for at least 18 months under these conditions.
The HiPure FastFilter Plasmid DNA Maxi 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 a special 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 1000 µg high copy number plasmid DNA or 50-500 µg low copy number plasmid DNA can be purified from 200mL overnight culture.
Details
Specifications
Features
Specifications
Main Functions
Isolation up to 1mg plasmid DNA from 200ml bacterial culture
Applications
Enzyme digestion, sequencing, PCR and labeling, etc.
Purification method
Maxi spin column
Purification technology
Silica technology
Process method
Manual (centrifugation or vacuum)
Sample type
Conventional plasmid, plasmid less than 30KB
Sample amount
100-200ml
Yield
0.4-1mg
Elution volume
≥500μl
Time per run
≤60 minutes
Liquid carrying volume per column
20ml
Binding yield of column
1mg
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: Preparation and 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
Economy – high cost performance
Kit Contents
Contents
P101402
P101403
Purification Times
10 Preps
50 Preps
RNase A
20 mg
40 mg
Buffer P1
140 ml
2 x 350 ml
Buffer P2
140 ml
2 x 350 ml
Buffer LEN3
70 ml
350 ml
Buffer GBT
120 ml
550 ml
Buffer PW1
60 ml
300 ml
Buffer PW2*
50 ml
4 x 100 ml
Elution Buffer
20 ml
120 ml
HiPure DNA Maxi Columns III
10
50
Lysate Clear Maxi Syringe
10
50
50 ml Collection Tubes
20
100
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 Maxi 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 a special 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 1000 µg high copy number plasmid DNA or 50-500 µg low copy number plasmid DNA can be purified from 200mL overnight culture.
Solid Phase Reversible Immobilization magnetic beads consist of paramagnetic particles coated with carboxyl groups that reversibly bind DNA. They are used for DNA purification because they are fast, simple and efficient. We have developed our own beads technology that are different from other SPRI beads technologies.
Our Magnetic Beads(DNA & RNA Purification) combines BioDynami’s proprietary chemistries with reversible DNA-binding properties of magnetic beads. The magnetic beads, which are unique from other SPRI beads, are developed for effective nucleic acid purification by removing unwanted components such as salts, dNTPs, enzymes, primers, adapters, and other impurities. The beads are RNase free, can be used for applications of DNA, and even work with more sensitive RNA without any additional cost.
Our magnetic beads are optimized to selectively bind DNA fragments of 100 bp and larger, and RNA fragments of 200 bases and larger, similar to other SPRI beads such as AMPure® XP* and SPRIselect*. Purified DNA and RNA are suitable for downstream applications requiring high quality DNA and RNA, as the purified fragments are free of contaminants and impurities. The beads can be used for NGS library purification, PCR fragment cleanup, molecular cloning, or even nucleic acid concentration.
The beads can also be used for size selection of DNA fragments ranging from 150 bp to 800 bp by changing the bead-to-sample volume ratio and performing single or double-size selection. The beads are an ideal choice for NGS library preparation. They can be easily integrated into the standard workflow of NGS library preparation since the volume ratio is similar for protocols using standard magnetic beads.
Features:
Effective recovery of DNA and RNA samples
DNA fragments greater than 100 base pairs
RNA fragments greater than 200 bases
Removal of unwanted components and impurities
Fragment size selection for specific applications
Consistent single or double-size selection
Flexibility: compatible with manual and automated processing
Cost effective alternative to other beads such as AMPure® XP* and SPRIselect* with equivalent performance
* AMPure® XP and SPRIselect are trade marks of Beckman Coulter.
Magnetic beads recovery rate. dsDNA and ssDNA of genomic DNA, 1 kb DNA, and 200 bp DNA fragments were used. Total RNA was also tested.
Comparison of elution volume vs yield. 40 ul , 30 ul, and 20 ul of elution volume were used. BioDynami magnetic beads have better recovery rates at low elution volume when compared to other SPRI beads such as AMPure® XP*.
