High Pure PCR Cleanup Micro Kit
ROCHE/HPPCRCURO - sufficient for 50 purifications (04983955001), sufficient for 200 purifications (04983912001), suitable for DNA purification
Synonym: PCR purification
Product Type: Chemical
HP Cleanup PCR Application
Choose the right product for DNA fragment purification.
Figure 1: 1% agarose gel electrophoresis of 341 bp PCR product recovered in the presence of different amounts of Binding Enhancer. 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. Lanes 1: Molecular weight marker VI 2: 0% Binding Enhancer 3: 10% Binding Enhancer 4: 20% Binding Enhancer 5: 40% Binding Enhancer 6: PCR without purification 7: PCR negative control (PCR without template) 8: Molecular weight marker VI
Table 1: Yield of DNA fragments isolated in the presence of increasing amounts of Binding Enhancer. Increasing the amount of Binding Enhancer in the isolation procedure increases the recovery of DNA fragments from the PCR product mix. As shown in lanes 2–5, however, this increase mainly reflects the differing amounts of smaller PCR fragments (e.g., primer-dimers) co-purified with the PCR product.
Typical DNA Recovery
Principle
| manufacturer/tradename | Roche |
| packaging | pkg of 200 purifications (04983912001) |
| pkg of 50 purifications (04983955001) | |
| Quality Level | 100  |
| technique(s) | DNA purification: suitable |
| usage | sufficient for 200 purifications (04983912001) |
| sufficient for 50 purifications (04983955001) |
| 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. |
| Application: | 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. |
| Components: | • Binding Buffer • Binding Enhancer • Wash Buffer Concentrate • Elution Buffer • High Pure Micro Filter Tubes • Collection Tubes |
| 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 kit’s Binding Enhancer to adjust purification stringency. • Eliminate the use of hazardous organic compounds such as cesium chloride, phenol, chloroform, and ethidium bromide |
| General description: | 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 kit’s 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. |
| General description: | 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. |
| Legal Information: | LightCycler is a registered trademark of Roche |
| Other Notes: | High Pure Technology and Silica Adsorption Kits |
| Other Notes: | For life science research only. Not for use in diagnostic procedures. |
| 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. |
| Quality: | Greater than 70% recovery is obtained when 3 μg Roche DNA Molecular Weight Marker VI is applied to the kit’s 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. |
| Symbol |    GHS05,GHS07,GHS08,GHS05,GHS07,GHS08 |
| Signal word | Danger-Danger |
| Hazard statements | H302 + H332 - H314 - H360Df - H412 - H302 + H332 - H314 - H360FD - H412 |
| Precautionary statements | P201 - P280 - 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 |
| Supplemental Hazard Statements | EUH019 - EUH032 |
| UNSPSC | 41105500 |