Tri(propargyl-PEG5-NHCO-ethyloxyethyl)amine is a click chemistry branched linker. The propargyl groups can react with azide-bearing molecules via copper catalyzed Click Chemistry.

Tri(propargyl-PEG5-NHCO-ethyloxyethyl)amine is a click chemistry branched linker. The propargyl groups can react with azide-bearing molecules via copper catalyzed Click Chemistry.

Introduction

The HiPure Midi system provides a fast, simple, and cost-effective plasmid DNA midiprep method for routine molecular biology laboratory applications. HiPure Midiprep Kits use silica membrane technology to eliminate the cumbersome steps associated with loose resins or slurries. Plasmid DNA purified with Mini Kits is immediately ready for use. Phenol extraction and ethanol precipitation are not required, and high quality plasmid DNA is eluted in a small volume of Tris buffer or water.

Details

Specifications

Features	Specifications
Main Functions	Isolation up to 500µg endotoxin-free plasmid DNA from 25-50ml bacterial culture. Recommend for High copy vector, Low elution volume, High concentration
Applications	Cell transfection, animal injection, etc.
Purification method	Midi spin column
Purification technology	Silica technology
Process method	Manual (centrifugation or vacuum)
Sample type	High copy plasmid vector
Sample amount	25-50ml LB
Yield	10-500μg
Elution volume	≥0.1ml
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: 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 concentration – easily obtain concentrations >1000ng/μl
• Small amount large extraction – using a small amount of column to achieve a large extraction yield (500μg)
• Low endotoxin content – the obtained plasmid can be directly used in cell transfection and animal injection

Kit Contents

Contents	P123102	P123102B
Purification Times	50 Preps	50 Preps
RNase A	30 mg	30 mg
Buffer E1	150 ml	150 ml
Buffer E2	150 ml	150 ml
Buffer E3	150 ml	150 ml
Buffer E4	150 ml	150 ml
Buffer E5	150 ml	150 ml
Buffer PW2*	50 ml	50 ml
Elution Buffer	30 ml	30 ml
MaxPure EF Mini Column	50	50
2 ml Collection Tubes	50	50
Lysate Clear Midi Syringe	50	50
Extend Tubes	50	50
50ml Centrifuge Tubes		50
Support Tubes		50

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 and optional LyseBlue reagent, 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 Midi system provides a fast, simple, and cost-effective plasmid DNA midiprep method for routine molecular biology laboratory applications. HiPure Midiprep Kits use silica membrane technology to eliminate the cumbersome steps associated with loose resins or slurries. Plasmid DNA purified with Mini Kits is immediately ready for use. Phenol extraction and ethanol precipitation are not required, and high quality plasmid DNA is eluted in a small volume of Tris buffer or water.

The NGS FFPE DNA Library Prep Kit (illumina and MGI Platforms) was developed for the construction of high quality FFPE DNA libraries using 10 ng to 400 ng of input DNA isolated from formalin-fixed, paraffin-embedded (FFPE) samples. The DNA damage caused by fixation makes it difficult to construct high quality libraries. Our kit has been optimized to repair damaged DNA in the reactions. Multiplexing of the NGS FFPE DNA Library is possible.

NGS FFPE DNA Library Prep Kit Workflow

FFPE samples are a great resource for biomedical research. However, the methods for fixation and condition of storage significantly damage the DNA in the samples.  Thus, the extraction of high quality DNA from FFPE samples is often a challenge.  Low yield and low quality of FFPE DNA usually are common because of the limited tissue material and the DNA degradation.

As a result, it is usually difficult to construct high quality NGS libraries from low amount and low quality of FFPE DNA. In order to address this issue, we have developed the NGS FFPE DNA Library Prep Kit to make high quality libraries from the low input of FFPE DNA samples.

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

Non-index (Cat.# 30035): Libraries do not have index.

Index (Cat.# 30037): Each of our index primers contains a unique barcode DNA (6 bases long) that can be used to identify individual library. Multiplexing of libraries is up to 48 samples. Index information can be downloaded here.

Unique dual index (Cat.# 30039): FFPE DNA library multiplexing is possible with 96 samples based on the unique dual indexing system. Our unique Four–Base Difference Index System allows us to make indexes that have at least 4 bases different from each other in the 8-base index sequence. Our unique dual indexing primers can effectively remove NGS errors including index hopping, de-multiplexing errors, index cross-contamination, mis-assignments etc. The unique dual index primer set includes 96 pre-mixed unique pairs of i5 and i7 index primers. Index information can be downloaded here.

Indexes are available for the MGI platform kits (Cat.# 34037).

Kit advantages:

• Fast and simple protocol
  • Making libraries in just 1.5 hours
  • 10 minutes of hands-on time
• Easy procedure
  • Ready-to-use master mix to reduce the tedious mixing
  • Less reaction components to simplify the reaction setup
• Less magnetic beads needed for the purification steps: saving more than 50% of the expensive beads
• Low input FFPE DNA: From 10 ng to 400 ng

Comparison of library conversion efficiency under the same condition. Input DNA amount is 25 ng.

Comparison of library yield under the same condition. Input DNA amount is 25 ng.

The NGS FFPE DNA Library Prep Kit (illumina and MGI Platforms) was developed for the construction of high quality FFPE DNA libraries using 10 ng to 400 ng of input DNA isolated from formalin-fixed, paraffin-embedded (FFPE) samples. The DNA damage caused by fixation makes it difficult to construct high quality libraries. Our kit has been optimized to repair damaged DNA in the reactions. Multiplexing of the NGS FFPE DNA Library is possible.