Introduction

The HiPure Plasmid EF Mini Kit combine the power of HiPure technology with Magen’s innovative endotoxin removal technology to deliver high-quality plasmid DNA with low endotoxin levels foruse in eukaryotic transfection, and in vitro experiments. The HiPure plasmid endo-free system uses a specially formulated buffer that prevents endotoxin molecules from binding to the surface of the HiPure matrix. Endotoxin contamination lowers transfection efficiencies for endotoxin sensitive celllines. For gene therapy, endotoxin contamination should be of major concernsince endotoxins have the potential to cause fever, endotoxin shock syndrome, and interfere with in vitro transfection into immune cells.

Details

Specifications

Features	Specifications
Main Functions	Isolation up to 80µg endotoxin-free plasmid DNA from 5-15ml bacterial culture. Recommend for low copy vector, Thoroughly remove RNA
Applications	Cell transfection, animal injection, etc.
Purification method	Mini spin column
Purification technology	Silica technology
Process method	Manual (centrifugation or vacuum)
Sample type	Low copy plasmid vector
Sample amount	5-15ml LB
Yield	10-70μg
Elution volume	≥75μl
Time per run	≤40 minutes
Liquid carrying volume per column	800μl
Binding yield of column	70μg

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 – silica gel column purification is much faster than ion exchange
• High yield – up to 70μg plasmid can be binded in one column
• Low endotoxin content – the obtained plasmid can be directly used in cell transfection and animal injection

Kit Contents

Contents	P115402	P115403
Purification Times	50 Preps	250 Preps
RNase A	5 mg	20 mg
Buffer P1	30 ml	140 ml
Buffer P2	30 ml	140 ml
Buffer LEN3	15 ml	70 ml
Buffer LN4	50 ml	250 ml
Buffer LN5	30 ml	140 ml
Buffer PW1	30 ml	140 ml
Buffer PW2	12 ml	50 ml
Elution Buffer	15 ml	30 ml
HiPure DNA Mini Columns III	50	250
2 ml Collection Tubes	50	250

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.

Purchase Guide

Name	CAT NO	Features	Sampleamount	Elutionvolume	Yield
HiPure Plasmid Mini Kit	P1001	Classic, Regular application	1-5ml LB	30-50μl	5–35µg/5ml
HiPure Plasmid Mini Kit II	P1002	Classic, Regular application	5-15ml LB	75-100μl	20-80µg/15ml
HiPure Plasmid Plus 96 Kit	P1006	Classic, Regular application	1-1.5ml LB	70-150μl	1-15μg/1ml
HiPure Fastfilter Plasmid Midi Kit	P1013	Fast	30-50ml LB	0.3-0.8ml	50–250µg/50ml
HiPure Fastfilter Plasmid Maxi Kit	P1014	Fast	100-200ml LB	0.5-1.5ml	0.4–1mg/200ml
Low endotoxin/Endotoxin-free Plasmid
HiPure Plasmid EF Maxi Kit	P1156	Classic method, General for all kinds of vectors	100-200ml LB	≥0.7ml	200–1500µg
HiPure Plasmid EF Mini Kit	P1154	Recommend for low copy vector, Thoroughly remove RNA	5-15ml LB	≥60μl	10–80µg
HiPure Plasmid EF 96 Kit	P1157	Recommend for low copy vector, Thoroughly remove RNA	1-5ml LBx96	≥75μl	30µg
HiPure Plasmid EF Midi Kit	P1231	Recommend for High copy vector, Low elution volume, High concentration	25-50ml LB	≥100μl	10-500μg
HiPure Plasmid EF Mega Kit	P1116	Recommend for High copy vector(>3μg/ml)	500ml LB	≥2ml	1-10mg

For any technical problems or customized products, please contact us.

F&Q about Endotoxin-free Plasmid Extraction Kit — P1156 ←click here

The HiPure Plasmid EF Mini Kit combine the power of HiPure technology with Magen’s innovative endotoxin removal technology to deliver high-quality plasmid DNA with low endotoxin levels foruse in eukaryotic transfection, and in vitro experiments. The HiPure plasmid endo-free system uses a specially formulated buffer that prevents endotoxin molecules from binding to the surface of the HiPure matrix. Endotoxin contamination lowers transfection efficiencies for endotoxin sensitive celllines. For gene therapy, endotoxin contamination should be of major concernsince endotoxins have the potential to cause fever, endotoxin shock syndrome, and interfere with in vitro transfection into immune cells.

DBCO-PEG5-DBCO is a bifunctional click chemistry reagent. DBCO will react with azide-bearing compounds or biomolecules to form a stable triazole linkage without copper catalyst. The hydrophilic PEG spacer arm can increase water solubility and membrane permability. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

DBCO-PEG5-DBCO is a bifunctional click chemistry reagent. DBCO will react with azide-bearing compounds or biomolecules to form a stable triazole linkage without copper catalyst. The hydrophilic PEG spacer arm can increase water solubility and membrane permability. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

The NGS Low Input DNA Library Prep Kit (illumina and MGI Platforms) was developed for construction of high quality NGS libraries with low input DNA amount from 1 ng to 400 ng. The kit allows scientist to study samples with limited DNA such as tumor samples, patient samples, and other specially collected samples (FACS sorting etc.). The kit has high library conversion efficiency with as little as 1 ng DNA input. The fast and simple 1.5-hour protocol makes libraries with even coverage and low GC-bias based our unique chemistry for DNA end-polishing and ligation.

The kit needs double strand DNA fragments (blunt and/or sticky) as input DNA for NGS library preparation, and is compatible with DNA fragments generated from both enzymatic approach (for example, BioDynami DNA fragmentation enzymes, Cat.# 40061 and Cat.# 40062) and mechanical approaches such as sonication and nebulization etc.

NGS Low Input DNA Library Prep Kit Workflow

Three index types are available for the NGS Low Input DNA Library Prep Kit of illumina platform:

Non-index (Cat.# 30022): Libraries do not have index.

Index (Cat.# 30024): Each of our index primers contains a unique barcode sequence with 6 bases that can be used to identify the low input DNA libraries. Library multiplexing up to 48 samples is possible. Index information can be downloaded here.

Unique dual index (Cat.# 30025): Library multiplexing up to 96 low input DNA libraries is possible with unique dual indexes with our Four-Base Difference Index System. The system allows us to make indexes for the libraries that have at least 4 bases different from each other in the 8-base barcode length. Our unique dual index primers can reduce sequencing errors such as de-multiplexing errors, amplification errors, mis-assignment of reads, index cross-contamination, and also index hopping. The kit  includes 96 pre-mixed unique pairs of index primers. Index information can be downloaded here.

Indexes are available for the MGI platform kits (Cat.# 34024).

Kit advantages:

• • • Fast protocol
      • The hands-on time is only 10 minutes
      • The total protocol time is around 1.5 hours
    • Simple procedure
      • Ready-to-use master mix makes it simple for reaction setup
      • Less reaction components
    • Less magnetic beads required for cleanup steps: Save the cost more than 50%
    • Low input DNA amount: Starts from 1 ng of DNA

Comparison of library conversion efficiency under the same NGS library preparation condition. Input DNA amounts are 1 ng and 10 ng, respectively. DNA was mechanical sheared with Covaris before library prep. BioDynami kit: NGS Low Input DNA Library Prep Kit (Cat. #30022).

Comparison of library yield under the same NGS library prep condition. Input DNA amounts are 1 ng, 10 ng, and 100 ng. DNA was mechanical sheared with Covaris before library prep. BioDynami kit (Cat. #30022). PCR cycle numbers were indicated.

The NGS Low Input DNA Library Prep Kit (illumina and MGI Platforms) was developed for construction of high quality NGS libraries with low input DNA amount from 1 ng to 400 ng. The kit allows scientist to study samples with limited DNA such as tumor samples, patient samples, and other specially collected samples (FACS sorting etc.). The kit has high library conversion efficiency with as little as 1 ng DNA input. The fast and simple 1.5-hour protocol makes libraries with even coverage and low GC-bias based our unique chemistry for DNA end-polishing and ligation.