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| 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.
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- 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
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| - Sequencing, cloning, ligation, probe labeling
ligation, random primed labeling, nick translation etc.
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| * 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. |
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 Purification
yield |
| 1. % Recovery of DNA from the different fragment length
& amount of DNA |
[ Table
1. the different fragment length ]
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[ Table
2. the amount of DNA ] |
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Fragment length |
% Recovery |
| 50bp
- 200bp |
80% |
| 200bp
- 4Kb |
95% |
| 4Kb - 10Kb |
90% |
| > 10 Kb |
75% |
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Amount of DNA |
% Recovery |
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5 - 10ug |
80% |
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10 - 30ug |
95% |
| > 30ug |
75% |
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| 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 |
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| 3. Comparison of DNA recovery with various starting PCR
product volume |
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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 |
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| Purification
purity |
1.
Enzyme digestion result |
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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 |
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| Comparison
of DNA yield from different company kit |
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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 |
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