High Pure PCR Cleanup Micro Kit

Кат. номер

4983955001

4983912001

Nucleic acids bind to the surface of the glass fiber fleece in the presence of a chaotropic salt (guanidine HCl). This allows the High Pure filter tube to specifically immobilize nucleic acids (both DNA and RNA) while they are freed of contaminants. The amount of DNA recovered is dependent on the amount of DNA applied to the glass fiber fleece, the elution volume, and the length of the amplification/DNA products. When 5 - 25 µg DNA is applied to the kits High Pure Micro Filter Tube, approximately 85% of the DNA can be recovered.
Capacity of High Pure Micro Filter Tubes: The High Pure Micro Filter Tubes hold up to 500 µL sample volume.
The High Pure PCR Cleanup Micro Kit efficiently purifies products from PCR and other reactions. The kit eliminates primers, mineral oil, salts, unincorporated nucleotides, and thermostable DNA polymerases, which may inhibit subsequent enzymatic reactions such as labeling, sequencing, or cloning of PCR products.
The High Pure PCR Cleanup Micro Kit is used to isolate PCR products from amplification and other reactions, and can be used in many molecular biology applications:

• Restriction enzyme digests
• Alkaline phosphatase treatment
• Kinase reactions
• Nonradioactive labeling

The kit can also be used to:

• Purify cDNA
• Concentrate dilute nucleic acid solutions
• Recover DNA from agarose gel slices

Use one kit for a variety of applications.
The fast and simple High Pure protocols use a tabletop centrifuge to bind, wash, and elute the reaction product down to 10 µL (micro format) in as little as 10 minutes. The procedure conveniently eliminates a concentration step, and is ideal for downstream applications such as labeling, sequencing, cloning, ligation, or amplification using PCR. The purified DNA can also be used for Southern blotting and in vitro transcription.
Features and Benefits

• Conserve resources and save time

by using a single kit with a simple, rapid protocol.

• Obtain purified product in a small elution volume

(10 µL) for demanding downstream applications.

• Generate contaminant-free DNA

for direct use in cloning, ligation, restriction digests, and other reactions.

• Selectively isolate specific DNA fragment sizes

by using the kits Binding Enhancer to adjust purification stringency.

• Eliminate the use of hazardous organic compounds

such as cesium chloride, phenol, chloroform, and ethidium bromide
Components

• Binding Buffer
• Binding Enhancer
• Wash Buffer Concentrate
• Elution Buffer
• High Pure Micro Filter Tubes
• Collection Tubes

Quality
Greater than 70% recovery is obtained when 3 µg Roche DNA Molecular Weight Marker VI is applied to the kits High Pure Micro Filter Tubes. Gel electrophoresis of the eluate after purification confirms the complete removal of primer-dimers. The eluted, purified DNA shows no inhibition of amplification using a LightCycler® Instrument with the LightCycler® FastStart DNA MasterPLUS SYBR Green I.
Preparation Note
Nucleic acids bind specifically to the surface of glass fibers in the presence of chaotropic salts. Since the binding process is specific for nucleic acid, the bound material can be separated and purified from impurities by a simple wash step. The Binding Enhancer enables the modification of DNA fragment size exclusions. Small oligonucleotides and dimerized primers from amplification reactions are selectively removed. The nucleic acids elute from the glass fiber fleece in a low-salt buffer or water.
Analysis Note
A 341 bp PCR fragment of the tPA gene was amplified according to a standard block cycler protocol. The resulting reaction mixes were pooled and purified with the High Pure PCR Cleanup Micro Kit. Different amounts of Binding Enhancer were used in the purification procedure. Portions of the PCR product (250 ng each, lanes 2 – 5) and the PCR product mix (16 µL each, lanes 6 – 7) were analyzed on a 1% agarose gel (see Figure 1).
The yield from each purification is shown in Table 1.
High Pure Technology and Silica Adsorption Kits,
For life science research only. Not for use in diagnostic procedures.
LightCycler is a registered trademark of Roche

Quality Level	100,
Торговая марка	Roche
packaging	pkg of 200 purifications (04983912001), pkg of 50 purifications (04983955001)

pictograms	GHS05,GHS07,GHS08
signalword	Danger
hcodes	H302 + H332 - H314 - H360Df - H412
pcodes	P201 - P280 - P303 + P361 + P353 - P304 + P340 + P310 - P305 + P351 + P338 + P310 - P308 + P313
RIDADR	NONH for all modes of transport
WGK Germany	WGK 2
Flash Point F	285.8 °F
Flash Point C	141 °C