This kit provides a rapid spin column procedure for the purification and clean-up of sequencing and various other enzymatic reactions including restriction enzyme digests, Klenow reactions, alkaline phosphatase reactions, and ligations. The kit is used to remove reaction contaminants including dye terminators, salts, enzymes, excess primers and primer dimers. Contaminants are undesirable as they can interfere with many downstream applications including sequencing, RFLP, restriction enzyme digestions and ligation. Purification is based on spin-column chromatography without the use of phenol, chloroform or alcohol precipitation. The kit provides a high quality product with up to 90% recovery.
The kit is also available in a 96-well format for high-throughput sequencing reaction clean-up. Purification with the 96-well plate can be performed using either a vacuum manifold or centrifugation.
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| Kit Specifications – Spin Column | |
| Column Binding Capacity | 13 μg |
| Maximum Column Loading Volume | 650 μL |
| Maximum Amount of Starting Material | 25 μg |
| Time to Complete 10 Purifications | 10 minutes |
| Minimum Elution Volume | 30 μL |
Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 1 year after the date of shipment.
| Component | Cat. 34500 (50 preps) | Cat. 34400 (2 x 96-well plate) |
|---|---|---|
| Binding Buffer C | 30 mL | 3 x 30 mL |
| Wash Solution A | 12 mL | 2 x 20 mL |
| Elution Buffer B | 8 mL | 2 x 15 mL |
| Spin Columns | 50 | – |
| Collection Tubes | 50 | – |
| Elution Tubes (1.7 mL) | 50 | – |
| 96-Well Plate | – | 2 |
| Adhesive Tape | – | 4 |
| 96-Well Collection Plate | – | 2 |
| 96-Well Elution Plate | – | 2 |
| Product Insert | 1 | 1 |
HiDi is available as:
HiDi® DNA Polymerase (>>)
HiDi® Taq DNA Polymerase (>>)
HiDi® 2x PCR Master Mix (>>)
HiDi® Taq 2x PCR Master Mix (>>)
Casestudies:
HiDi® DNA Polymerase: Applications from mutation detection to genome editing (read more)
Matching vs. mismatching nucleotide is placed at the 3′-end of the primer for best discrimination results.
Cilantro: some people love it in their food, some hate it. Here we are detecting a genomic SNP (rs72921001) in HeLa genomic DNA. This SNP is reported to be close to a number of genes coding for olfactory receptors. (Reference: Eriksson N. et al. (2012), “A genetic variant near olfactory receptor genes influences cilantro preference.”)
Considering, that only the C-allele specific primer is extended and yielding in a specific amplicon, we can conclude a genetic predisposition in disliking cilantro, as this SNP is significantly associated with detecting a soapy taste to cilantro.
Allele-specific PCRs were performed from 1 ng/µl of HeLa gDNA in the presence of a realtime dye, indicating the amplification of the C-allele specific primer only. The A-allele specific primer is discriminated, thus not amplified up to 50 cycles.
PCR products were subsequently analysed on a 2.5% agarose gel. Specific product is visualized by ethidium bromide staining at the amplicon length of 109 bp.
HiDi® stands for High Discrimination of mismatches at the 3’-terminus of primers in PCR. This myPOLS Biotec enzyme family is optimized for this feature and is the first choice for applications that rely on this property such as allele-specific PCR (asPCR) that is also termed allele-specific amplification (ASA).
m-PEG12-DBCO is a long chain click chemistry PEG reagents. The DBCO group can react with azides via a copper-free Click Chemistry reaction to form a stable triazole. This reagent can be applied to situations when copper is a concern for DBCO-activated PEGylation. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
m-PEG12-DBCO is a long chain click chemistry PEG reagents. The DBCO group can react with azides via a copper-free Click Chemistry reaction to form a stable triazole. This reagent can be applied to situations when copper is a concern for DBCO-activated PEGylation. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.