<D6915> Endo-Free Plasmid DNA Midi Kit 无内毒素质粒中量提取试剂盒

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无内毒素质粒中量提取试剂盒

Endo-Free Plasmid DNA Midi Kit

产品编号	产品名称	包装规格	目录价
D6915-01	无内毒素质粒中量提取试剂盒	10T	630
D6915-03	无内毒素质粒中量提取试剂盒	25T	1530
D6915-04	无内毒素质粒中量提取试剂盒	100T	5670

产品介绍

The E.Z.N.A.® Endo-Free Plasmid DNA Midi Kit provides an efficient method for the large-scale isolation of low-endotoxin plasmid DNA. The system utilizes Lysate Clearance Filter Syringes instead of centrifugation to rapidly clear bacterial lysates post-alkaline lysis. Following lysate clearance, endotoxins are removed, and the protocol follows a simple “bind-wash-elute” procedure to deliver high-quality plasmid DNA. The yields vary according to plasmid copy number, E. coli strain, and growth conditions. 15-50 mL of bacterial cultures in LB medium typically produces up to 200 µg of high copy number plasmid DNA. Up to 100 mL of culture may be processed when working with low copy number plasmid. The system uses centrifugation or vacuum technology for plasmid purification and eliminates the time-consuming gravity-flow columns that require alcohol precipitation. Purified plasmid DNA is suitable for sensitive downstream applications such as transfection of mammalian cells, cloning, automated fluorescent DNA sequencing (typical reads exceed 800 bp), next-generation sequencing, restriction enzyme digestion, ligation, PCR, in vitro transcription, transformation, etc.

参数

FEATURES	SPECIFICATIONS
Downstream Application	Transfection, gene therapy, cloning, in vitro Transcription, Next-Generation Sequencing, PCR, Transformation, restriction digestion, ligation
Starting material	15–50 mL LB culture with OD600 between 2 and 3; or equivalent
Plasmid type	High-copy, low-copy, cosmid DNA
Processing mode	Manual (centrifugation or vacuum)
Throughput	1 – 24
DNA binding technology	Silica Midi Spin Column
Lysate clearance method	Filtration using Lysate clearance syringe
Processing time	45 minutes
Yield	100–250 μg for high copy-number; 10–50 μg for low copy-number
Special note	Includes endotoxin removal step

组分

Endo-Free Plasmid DNA Midi Kit	D6915-00	D6915-01	D6915-03	D6915-04
Purifications	2	10	25	100
HiBind® DNA Midi Columns	2	10	25	100
15 mL Collection Tubes	2	10	25	100
Lysate Clearance Filter Syringe	2	10	25	100
Solution I	6 mL	30 mL	80 mL	270 mL
Solution II	6 mL	30 mL	80 mL	270 mL
N3 Buffer	3 mL	15 mL	40 mL	140 mL
ETR Binding Buffer	15 mL	60 mL	170 mL	650 mL
ETR Wash Buffer	6 mL	30 mL	60 mL	200 mL
EHBC Buffer	4 mL	21 mL	48 mL	200 mL
DNA Wash Buffer	5 mL	25 mL	50 mL	200 mL
Endo-Free Elution Buffer	15 mL	30 mL	60 mL	250 mL
RNase A	20 μL	100 μL	300 μL	1.2 mL
User Manual	✓	✓	✓	✓

流程

Centrifugation Protocol

Pellet by Centrifugation
Resuspend, Lyse, Neutralize
Transfer Lysate to Syringe
Insert Plunger into Syringe and Clear Lysate into 15 mL Collection Tube
Add ETR Binding Buffer
Pass Lysate through HiBind® DNA Column
Wash 4X
Dry HiBind® matrix
Elute Endotoxin-free DNA

Vacuum Protocol

Pellet by Centrifugation
Resuspend, Lyse, Neutralize
Transfer Lysate to Syringe
Insert Plunger into Syringe and Clear Lysate into 15 mL Collection Tube
Prepare Vacuum Manifold
Add ETR Binding Buffer
Insert column into leur connector on vacuum manifold
Pass Lysate through HiBind® DNA Column
Wash 4X
Dry HiBind® matrix
Elute Endotoxin-free DNA

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