High Pure PCR Template Preparation Kit
The High Pure PCR Template Preparation Kit efficiently isolates nucleic acids from a diverse array of sample materials such as whole blood, cultured cells, and tissues, preparing them for direct use in PCR and restriction digest reactions. Specifically designed to handle bacteria and yeast with prelysis treatments like lysozyme or lyticase, this kit facilitates the extraction of nucleic acids ready for critical molecular biology applications. Incorporating key components like Proteinase K for cell lysis, glass fibers for nucleic acid binding, and an Inhibitor Removal Buffer to eliminate PCR inhibitors, it streamlines the purification process to produce high-quality DNA and RNA. This kit represents an ideal solution for achieving accurate PCR amplifications and is especially suitable for applications requiring long template or high molecular weight DNA isolation.
Benefits of the High Pure PCR Template Preparation Kit
- Maximize flexibility in sample preparation.
- Works well with a wide variety of sample materials.
- Enhance amplification results.
- Efficiently remove PCR inhibitors.
- Facilitate long template applications.
- Isolate high molecular weight DNA.
- Improve PCR accuracy.
- Reproducibly obtain high-quality DNA, even in real-time PCR.
How the High Pure PCR Template Preparation Kit works
Cells are lysed using a short incubation with Proteinase K in the presence of a chaotropic salt (guanidine-HCl). Cellular nucleic acids (NA) bind selectively to special glass fibers pre-packed in the High Pure Purification Filter Tube. Bound nucleic acid is purified in a series of rapid “wash-and-spin” steps to remove contaminating cellular components. A specially formulated Inhibitor Removal Buffer has been included for use with sample material treated with 100 U/ml of heparin. Low-salt elution is used to release nucleic acid from the glass fiber. This simple method eliminates the need for organic solvent extractions and DNA precipitation, ideal for rapidly purifying many samples simultaneously.
- Blood, cells, or tissue are lysed by incubation with a special Lysis Buffer and Proteinase K.
- Nucleic acids are bound to the glass fibers pre-packed in the High Pure Filter Tube.
- Bound nucleic acids are washed with a special Inhibitor Removal Buffer to get rid of PCR inhibitory contaminants.
- Washing of bound nucleic acids, purification from salts, proteins, and other cellular impurities.
- Purified nucleic acids are recovered using the Elution Buffer.
Table: Range of sample volume and concentration, as well as expected results. Each Isolation was performed with 12 replicates followed by duplicate measurements on the LightCycler ® 480 Instrument II to assess the mean C
