Bioclone/BcMag™ One-Step Bacteria DNA Purification Kit/100x preps/AE102

Standard procedures for extracting bacteria DNA can be exceedingly Time consuming and labor intensive—specific applications like bacteria genotyping demand a lengthy DNA extraction method. For example, traditional bacteria DNA extractions need a 4-hour to overnight digestion with proteinase K, followed by phenol: chloroform extraction and alcohol precipitation. Other techniques, such as silica bind and elute kits, have recently become available. Those techniques are based on positive chromatography purifying selection. High chaotropic salt concentrations, such as guanidine hydrochloride, bind DNA to silica. The silica column or beads is washed with a salt/ethanol solution after nucleic acid binding to eliminate additional biomolecules from the sample. Finally, the column or beads is eluted using Tris elution buffer or water to remove the pure DNA. Such bind-wash-elute procedures are time-consuming, requiring multiple washing and spinning steps. Repetitive spin steps can cause considerable DNA loss (20-40%) and shearing. Furthermore, chaotropic salts and other impurities can easily pass through the eluted DNA or RNA, compromising ultimate purity and quantification as well as downstream enzymatic activities like PCR.

BcMag™ One-Step Bacteria DNA Purification Kit allows rapid, column-free extraction of genomic DNA from microorganisms, such as Gram-positive or Gram-negative bacteria. The kit uses our unique proprietary magnetic beads and buffers to efficiently lyse cells and remove all impurities simultaneously in an aqueous buffer, leaving the DNA untouched. The procedure employs mild lysis conditions, avoiding harsh conditions such as alkaline lysis and toxic chemicals for lysing cells to maintain DNA integrity and the time-consuming cleanup of organic solvent from the sample. Furthermore, the magnetic beads eliminate PCR inhibitors (Fig.1) from samples in a single step without DNA extraction. It increases DNA integrity, boosts nucleic acid yields, and minimizes DNA loss caused by typical DNA purification techniques’ time-consuming “bind-wash-elute” procedure. Following sample lysis, the straightforward one-step purification technique enables simultaneous processing of >96 samples and produces pure DNA in less than 30 minutes. Purified genomic DNA has the highest integrity and can be used in various downstream applications such as qPCR.

Workflow of BcMag™ One-Step Bacteria DNA Purification Kit

1.

Use bead beating to disrupt the sample in a bead beater.

2.

Centrifuge and transfer the supernatant to a new tube.

3.

Mix the samples with the magnetic beads and proteinase K and heat to lyse the cells.

4.

Vortex the beads to capture PCR inhibitors.

5.

Remove the beads with a magnet.

6.

Aspirate supernatant containing the pure ready-to-use DNA.

Performance

The purified DNA is ready for downstream applications, such as PCR, qPCR, single-nucleotide polymorphism (SNP), short tandem repeat (STR) genotyping, genotyping, next-generation sequencing (NGS, veterinary genotyping, etc.

Features and Advantages

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Rapid and efficient purification protocol: without prior DNA isolation for subsequent use in direct workflows, No liquid transfer, and One-tube.

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Ultrafast: Process 96 samples in less than an hour.

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Highest nucleic acids recovery rates: Minimal loss of DNA during extraction.

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Cost-effective: Eliminates columns, filters, laborious repeat pipetting, and organic reagents.

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High-throughput: Compatible with many different automated liquid handling systems.

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Instruction Manual

MSDS