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App. Package, M48, Forensics

For easy setup of protocols for forensics and genetic identity testing
  • More efficient yields from trace casework samples
  • Higher signal-to-noise ratios
  • “Tip dance” protocol for processing solid material
  • More optimized protocols, including large-volume protocol
  • Flexible elution volumes

The App. Package, M48, Forensics is a CD-ROM containing protocols for DNA purification from trace and reference forensic samples using the MagAttract DNA Mini M48 Kit on the BioRobot M48 workstation. These protocols provide operating commands for the workstation and allow easy setup through comprehensive, on-screen instructions.

Cat No./ID: 9016150
App. Package, M48, Forensics
Software protocol package for forensics applications, v2.1, on the BioRobot M48 workstation
The App. Package, M48, Forensics is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.
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Easy and efficient processing of solid materials with the “tip dance” protocol.
Papers from cigarette butts were spotted with 50 ng DNA per sample. After proteinase K digestion, solid materials were removed from half of the samples, which were then processed using the MagAttract DNA Mini M48 Kit with the standard trace protocol (Trace). The remaining half of the samples was processed using the "tip dance" protocol on the App. Package, M48, Forensics v2.0, without removing solid materials from the sample tubes (Tip dance). DNA yields were quantified by real-time, quantitative PCR.
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Higher yields of concentrated DNA with the large-volume protocol.
DNA was diluted in Buffer G2 to a final concentration of 50 pg/µl, and the indicated volumes were processed using the MagAttract DNA Mini M48 Kit with the standard trace protocol (Trace) or the large-volume protocol (LV) on the App. Package, M48, Forensics v2.0. All samples were eluted in 50 µl water, and 5 µl was quantified using real-time, quantitative PCR.
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Uniform yields with the normalization protocol.
DNA was purified from the indicated volumes of whole blood using the MagAttract DNA Mini M48 Kit with the standard trace protocol (Trace) or the normalization protocol (Normalization) on the App. Package, M48, Forensics v2.0. All samples were eluted in 50 µl water. DNA yields were uniformly limited to 150–250 ng using the normalization protocol.
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Higher signal-to-noise ratios with DNA purified from blood-stained fabric.
DNA was purified from dried blood (800 white blood cells) on cotton fabric and used for STR analysis. After proteinase K digestion, DNA was purified on the BioRobot M48 workstation using the MagAttract DNA Mini M48 Kit and [A] the v1.0 trace protocol on the App. Package, M48, Forensics v1.0 or [B]  the v2.0 trace protocol on the App. Package, M48, Forensics v2.0. PCR products were analyzed on an ABI PRISM 3100 Genetic Analyzer with Genotyper software. (Data kindly provided by M. Steinlechner, Institute of Legal Medicine, Innsbruck Medical University, Innsbruck, Austria.)
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High performance of DNA purified from saliva on cigarette butts.
DNA was purified from dried saliva (800 cells) on cigarette butts and used for STR analysis. After proteinase K digestion, DNA was purified on the BioRobot M48 workstation using the MagAttract DNA Mini M48 Kit and [A] the v1.0 trace protocol on the App. Package, M48, Forensics v1.0 or [B] the v2.0 trace protocol on the App. Package, M48, Forensics v2.0. PCR products were analyzed on an ABI PRISM 3100 Genetic Analyzer with Genotyper software.
(Data kindly provided by M. Steinlechner, Institute of Legal Medicine, Innsbruck Medical University, Innsbruck, Austria.)
Performance
Easier handling of solid materials with “tip dance” protocol

In addition to the standard trace protocol, an optional, fully automated “tip dance” protocol can be used, where the filter-tip moves back-and-forth relative to the worktable platform while pipetting. This enables processing of solid materials, such as swabs, fabrics, blood discs, or cigarette butts, directly in the sample tube. There is no need for prior centrifugation to remove solid materials that could clog the tip. The “tip dance” protocol simplifies handling and, in some cases, increases DNA yield with more efficient extraction of the sample (see figure "Easy and efficient processing of solid materials with the “tip dance” protocol").

Optimized protocol for larger sample volumes

A large-volume protocol enables fully automated processing of starting volumes up to 500 µl. This not only allows efficient DNA purification from dilute samples with low concentrations of DNA, such as diffuse stains, but also enables purification from samples that require larger volumes for thorough lysis. The ability to process larger sample volumes — with the same elution volumes as the standard trace protocol — enables higher yields of more concentrated DNA for greater sensitivity in downstream applications (see figure "Higher yields of concentrated DNA with the large-volume protocol").

 Normalization for uniform yields

Sample-to-sample variability can result in widely varying yields from reference blood or saliva samples. For routine, standardized analyses, purified DNA from these samples often needs to be quantified before analysis. Using the normalization protocol on the App. Package, M48, Forensics v2.1, DNA yields can be limited uniformly to 150–250 ng (see figure "Uniform yields with the normalization protocol"). This enables subsequent genetic analysis without the need to measure or adjust DNA concentration. The purified DNA is ready to use in downstream applications.

Flexible elution volumes

Purified DNA can be eluted in volumes from 50 to 400 µl (50 µl, 75 µl, 100 µl, 150 µl, 200 µl, 250 µl, 300 µl, or 400 µl), and is ready to use at concentrations needed for different downstream applications. DNA can be eluted in water or, at the user’s choice, in TE buffer for increased stability of small amounts of purified DNA.

Principle

The BioRobot M48 and the MagAttract DNA Mini M48 Kit, with protocols on the App. Package, M48, Forensics v2.1, reproducibly automate purification of genomic DNA from forensic, human-identity, and biosecurity applications. Purification of 6–48 samples per run is efficient and purified DNA performs well in downstream analyses, such as quantitative PCR and STR analysis, with high signal-to-noise ratios.

The BioRobot M48 system provides fully automated purification of DNA using silica-coated magnetic particles, eliminating the need for tedious centrifugation steps. The easy-to-use workstation allows purification of 6–48 samples in a single run. Between processing runs, the worktable can be decontaminated using an integrated UV lamp to reduce the chances of environmental contamination.

The MagAttract DNA Mini M48 Kit, with protocols on the App. Package, M48, Forensics v2.1, provides optimized chemistries for automated DNA purification with more efficient yields. STR analysis shows higher peaks for more sensitive detection (see figure "Higher signal-to-noise ratios with DNA purified from blood-stained fabric", note difference in y-axes). The higher peaks and improved signal-to-noise ratios enable highly sensitive detection and high performance in downstream applications (see figure "High performance of DNA purified from saliva on cigarette butts").

Procedure
The MagAttract DNA Mini M48 Forensic Handbook contains easy-to-use, optimized pretreatment protocols for specific types of forensic samples, including stains on fabric (see figure "Higher signal-to-noise ratios with DNA purified from blood-stained fabric"), cigarette butts (see figure "High performance of DNA purified from saliva on cigarette butts"), whole blood, swabs, chewing gum, bones, soil, and more. Separate protocols are included for setup of fully automated DNA purification on the BioRobot M48, including the standard trace protocol, a “tip dance” protocol, a large-volume protocol, and a normalization protocol.
Applications

The high-quality DNA is highly suited for many forensic, environmental, biodefense, biosecurity, and other applied-testing applications, such as:

Genotyping, including fingerprinting and paternity analysis
Purification of DNA from trace samples
DNA extraction from filters used in environmental testing
Routine analysis of reference samples
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