PCR Purification 96-Well Kit

Product Description

• Fast and easy processing - a 96-well plate can be processed in 20 minutes.
• Purification from all PCR by-products - remove all primer-dimers, primers, mineral oil, unicorporated nucleotides, enzymes and salts.
• High recovery - Recovery of greater than 90% of input amount.
• High Column Capacity: Bind up to 25 µg of PCR product
• Versatile - purify PCR products from 100 bp to 15 kb
• Can be used as an alternative to organic extraction and ethanol precipitation - this kit can also be used to clean-up various enzymatic reactions including digestions and Klenow reactions.
• Can also be used to remove PCR inhibitors (e.g. from milk DNA)
• Recovered DNA is suitable for all downstream applications - purified DNA is fully compatible with restriction digestions, ligations, labeling, PCR, and sequencing.
• If required, water can be used as elution buffer

The Norgen 96-well High Throughput PCR Purification Kit is designed for the rapid purification of PCR products from reaction mixtures.

The kit allows for the removal of mineral oil, enzymes, unincorporated nucleotides, primers and salts. Up to 95% of primer dimers are also removed with the kit. The recovered DNA is of a high quality, and is suitable for a number of downstream applications including restriction enzyme digestions, ligations, labeling, hybridizations and sequencing.

The kit employs the use of a novel and unique Silica Carbide Resin for DNA purification. This resin is an extremely inert and stable substance which is able to preferentially bind nucleic acids in the presence of high salts and low pH. Elution of the DNA is then dependent on a low salt concentration and higher pH conditions

An additional application of the Norgen PCR Purification Kit is that it can be used as an alternative to phenol extraction and ethanol precipitation in order to clean up various enzymatic reactions. Enzymes, buffers, dNTPs and other contaminants can be removed from the DNA using this kit as well.

Figure 1. High Yield Purification

The high yield of Norgen’s PCR Purification Kit is illustrated by purifying 3.5 µg of a 500 bp PCR-amplified product from a reaction mixture and comparing the recovery with a competitor. The purifications were performed in triplicate. DNA recovery was quantified by running 350 ng (1/10th) of the input and 5 µL (1/10th) of 50 µL DNA elution on a 1X TAE, 1% agarose gel followed by densitometry. The yield for each purification is shown in Figure 1 ((N1 to N3 for Norgen, and Y1 to Y3 for the competitor).

Norgen’s kit shows a significantly higher recovery than the competitor’s kit.

Figure 2. Removal of Primer-Dimers

Primer-dimers present in PCR reactions are effectively removed during the purification process with Norgen’s PCR Purification Kit. One microgram of a 700 bp PCR fragment, spiked with 250 pmole of primer (Lane 1) was purified using Norgen’s kit, with high recovery and no traces of spiked primer present in the elutions (Lanes 2 and 3). Lane 1 contains 400 ng (40%) of the input, and Lanes 2 and 3 contain 20 µL (40%) of the 50 µL elution (duplicate). Lane M is Norgen’s MidRanger 1kb DNA Ladder. Eluted DNA was resolved on a 1X TAE, 1% agarose gel.

Applications

DNA purified using Norgen's PCR Purification Kit can be used in a number of downstream applications including:

• Sequencing
• Ligations
• Labeling
• PCR
• Restriction Digestions

Kit Specifications

Binding Capacity Per Well: 15 ug

Maximum Loading Volume Per Well: 500 ul

Size of DNA Purified: 100 – 15,000 bp

Average DNA Recovery: > 90%

Average Primer Removal: > 90%

Minimum Elution Volume: 50 ul

Time to Complete 96 Purifications: 20 minutes

96-Well PCR Purification Kit Contents:

1. Binding Solution

2. Wash Solution

3. Elution Buffer

4. 96-Well Filter Plate

5. Adhesive Tape

6. 96-Well Collection Plate

7. 96-Well Elution Plate

8. Product Insert