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PCRquick-spin PCR Product Purification Kit

Ordering Information
Cat No Size
17201 50 col.
17202 250 col.


Price 17202

174.00 €

Add to Cart



Price 17201

77.00 €

Add to Cart

 

PROTOCOL

Description

The PCRquick-spin™ Kit procedure uses silica-membrane technology to remove nucleotides, dNTPs, enzymes, primers, mineral oil, salts, ethidium bromide, dyes, detergents and other impurities from PCR reactions quickly and efficiently. The PCRquick-spin™ products contain silica-gel membrane binding of up to 15ug DNA in high-salt buffer and eluting the DNA in low-salt buffer.

The system uses a simple bind-wash-elute procedure. PCR reaction samples are mixed with the appropriate binding buffer and then applied to the PCRquick-spin™ column where DNA binds to the silica-gel membrane. Impurities are washed away, and pure DNA is eluted in a small volume of the low-salt elution buffer provided or water, ready for use in any subsequent application.

Characteristics

- Takes only 15 minutes to extract DNA fragment
- Minimize DNA loss: recovers DNA fragment without solvent extraction, precipitation, or other steps that can lead to loss or degradation of DNA.
- High yield and excellent quality

Appilcation

- Sequencing, cloning, ligation, probe labeling ligation, random primed labeling, nick translation etc.

Kit Contents

Binding Buffer

150 ?

* Washing Buffer
: Add Absolute EtOH 200ml before use.

50 ?

* Elution Buffer

20 ?

* Columns
: Column contains membrane

250 columns

* Collection Tubes

250 Tubes

1) Optimized for 20-50? of PCR products, but could use upto 100?.
If more than 50? of PCR products is processed, add 700? of Binding buffer.
2) If the test is sensitive to salt, it is effective to proceed washing step twice.
3) It is suggested to use at least 20? of the Elution buffer to obtain the best result

ResultsTable 1. the different fragment length ]

Purification yield
1. % Recovery of DNA from the different fragment length & amount of DNA

 

Table 1. The different fragment lenght

Table 2. the amount of DNA

Fragment length

% Recovery
50bp - 200bp
80%
200bp - 4Kb
95%
4Kb - 10Kb
90%
> 10 Kb
75%

Amount of DNA

% Recovery

5 - 10ug

80%

10 - 30ug

95%

> 30ug

75%

 

2. Recovery of DNA from different PCR product size

Fig. DNA fragments extraction with the PCRquick-spinTM PCR Product Purification Kit

PCR product size : A, 131bp; B, 570bp; C, 830bp; D, 1.3Kb; E, 4.5Kb; F, 10Kb
Lane M, Maker DNA;
Lane 1,3,5,7,9,11, before purified with PCRquick-spinTM PCR Product Purification Kit;
Lane 2,4,6,8,10,12, after purified with PCRquick-spinTM PCR Product Purification Kit 

3. Comparison of DNA recovery with various starting PCR product volume

Fig. Comparison of DNA recovery with various starting PCR product volume
Lane M, Marker DNA;
Lane 1, 20? of PCR product;
Lane 2, 30? of PCR product;
Lane 3, 40? of PCR product;
Lane 4, 50? of PCR product;
Lane 5, 100? of PCR product

Purification purity
1. Enzyme digestion result

Fig. Enzyme digestion
3Kb PCR product was extracted using PCRquick-spinTM PCR Product Purification Kit and then digested with EcoRI. The digestions were performed at 37? for 1hr with each enzymes. And then result was analyzed ingel electrophoresis.
Lane M, Marker DNA;
Lane 1, 3, undigested PCR product, extracted using PCRquick-spinTM PCR Product Purification Kit;
Lane 2, 4, digested PCR product

Comparison of DNA yield from different company kit

Fig. Comparison of different company

Lane M, Marker DNA;
Lane 1, Control (not purify); Lane 2, iNtRON;
Lane 3, Supplier A; Lane 4, Supplier B

PCRquick-spin PCR Product Purification Kit
Ordering Information | Cat No 17202| Size 250 col.
174.00 €
Add to Cart

 

PCRquick-spin PCR Product Purification Kit
Ordering Information | Cat No 17201| Size 50 col.
77.00 €
Add to Cart

 
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