Protocol

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Contact us
nucleogen@hanmail.net for more information
Tel:031-315-6644     Fax:031-312-2008

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Spin column type

Ready-to-use high-purity plasmid DNA
(Mini : 1-10 ml, Midi : 10-100 ml, Maxi : 100-250 ml)

Just bind, wash, and elute

Economical kits

No alcohol precipitation, or organic extraction

Excellent reproducibility

Total procedure is 30 min

Recovery is 85-95% (Mini) or 80% - 90% (Midi and Maxi)

A260/280 ratio is > 1.8

Total procedure is 30 min

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

The Nucleogen Plasmid Purification Kit is designed for isolation of high-purity plasmid or cosmid DNA for use in routine molecular biology applications such as fluorescent and radioactive sequencing and cloning. Versatile Nucleogen Plasmid spin columns can be used either in microcentrifuges or on vacuum manifolds. The vacuum procedure provides simplified handling and faster sample processing. Nucleogen Plasmid spin columns can be vacuum processed using the commercial manifold with luer connectors.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 Mini (200preps)

for Negative Strain ……… 5112

for Positive Strain  ……… 7112

 Midi

10 preps  ……… 6101

50 preps  ……… 6105

100 preps  ……… 6110

 Maxi

6 preps  ……… 7106

24 preps  ……… 7124

50 preps  ……… 7150

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Principle

Nucleogen Plasmid spin columns contain a unique silica gel membrane that binds up to DNA in the presence of a high concentration of chaotropic salt, and allows elution in a small volume of low-salt buffer. Nucleogen Plasmid membrane technology eliminates time consuming phenol–chloroform extraction and alcohol precipitation, as well as the problems and inconvenience associated with loose resins and slurries. High-purity plasmid DNA eluted from Nucleogen Plasmid spin columns is immediately ready to use — there is no need to precipitate, concentrate, or desalt.


Procedure

Plasmid purification using Nucleogen Plasmid Purification Kits follows a simple bind–wash–elute procedure. First, bacterial cultures are lysed and the lysates are cleared by centrifugation. The cleared lysates are then applied to the Nucleogen Plasmid module where plasmid DNA adsorbs to the silicagel membrane. Impurities are washed away and pure DNA is eluted in a small volume of elution buffer or water.


Applications

Nucleogen Plasmid Purification Kits provide reproducible yields of high-purity DNA suitable for use in most applications, including:

Automated fluorescent sequencing, including capillary sequencing
Radioactive sequencing
Restriction digestion
Screening
Ligation and transformation
Labeling
PCR template preparation
All other exzymatic reactions

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

1   2   3   4   M  1   2   3   4

 

 

 

Plasmid DNA isolation using Nucleogen Plasmid Purification kit and Q company plasmid prep kit
Lane 1 : uncut pGEX-6p-1 plasmid DNA
Lane 2 : pGEX-6p-1 plasmid DNA digested Bam-HI
Lane 3 : uncut pQE 30-TAX plasmid DNA
Lane 4 : pQE 30-TAX plasmid DNA digested BamHI/HindIII
 

 

 

Kit Contents

 Mini

Mini-columns, Collection Tubes, Cell Resuspension Solution, Cell Lysis Solution, Neutralization Solution, Wash A Solution, Wash B Solution (7112), Elution Buffer, RNase A

 Midi

Midi-columns, Collection Tubes, Cell Resuspension Solution, Cell Lysis Solution, Neutralization Solution, Wash A Solution, Elution Buffer, RNase A

Maxi

Maxi-columns, Collection Tubes, Cell Resuspension Solution, Cell Lysis Solution, Neutralization Solution, Wash A Solution, Elution Buffer, RNase A