Bone DNA Extraction Kit
Rapidly Process Bone and Teeth Samples
- Quick preprocessing of bone powder using Demineralization Buffer
- Compatible with manual and Maxwell® DNA IQ™ chemistry
- Yields DNA free of inhibitors and compatible with downstream amplification reactions
- Note: The Bone DNA Extraction Kit (Cat.# DC6051) replaces the custom version of the Bone DNA Extraction Kit (Cat.# AX6780). No changes have been made to the product composition, and therefore, no performance changes are expected during the transition of the product from the custom to catalog version.
Catalog Number:
Size
Catalog Number: DC6051
Rapidly Process Challenging Skeletal Remains
Extraction of DNA from challenging bone samples. Femur bone samples dated from 1939–1945 (World War II) and 1870–1880 (Deadwood) were used with the Bone DNA Extraction Kit, Custom, followed by purification using DNA IQ™ chemistry on the Maxwell® Instrument. Autosomal DNA concentration, as determined by PowerQuant® System, was plotted as vertical bars. The normalized DNA or maximum template amount was used for amplification using the PowerPlex® Fusion System, and the number of alleles detected are shown.
In certain cases involving missing persons, mass disasters and anthropological excavations, bone and teeth are often the only sample types available for identification. Further, these samples are often subjected to environmental damage, and co-extraction of environmental inhibitors during purification limits their usability in STR typing experiments. Purification of DNA free of inhibitors is a crucial first step for obtaining a usable profile from these samples.
A short preprocessing protocol followed by purification of DNA using DNA IQ™ chemistry yields inhibitor-free DNA compatible with PowerQuant® or Plexor® HY Systems for human and male DNA quantification. This DNA can be used with the PowerPlex® STR Systems to generate an STR profile.
The article A Rapid and Efficient Method for DNA Extraction from Bone Powder demonstrates the use of Demineralization Buffer and subsequent purification of DNA using the Maxwell® FSC System with aged bone samples.
Promega forensic products are manufactured in alignment with the ISO 18385 standard.This standard ensures minimal risk of human DNA contamination for products used to collect, store and analyze biological materials for forensic purposes. Learn more.
Bone DNA Extraction Method Overview
In a 1.5ml microcentrifuge tube, 400µl of preprocessing buffer is added to bone powder and incubated at 56ºC for 2.5 hours on a shaker set at 1000rpm. The tube is then centrifuged for 5 minutes at 13,000 × g, and the supernatant is transferred to a new 1.5ml microcentrifuge tube containing Lysis Buffer with a reducing agent. At this point, the preprocessed sample is ready for purification using the DNA IQ™ manual or DNA IQ™ Maxwell® method. The 1.5ml microcentrifuge tube with the undigested bone powder pellets can be discarded. Following the purification protocol, DNA can be eluted in 50µl of elution buffer. The resultant DNA can be quantified using the PowerQuant® System and amplified using the PowerPlex® Systems to generate an STR profile.
Integration of the Bone DNA Extraction Kit into a laboratory's workflow offers a rapid, cost-effective means to extract DNA with minimal hands-on time from precious bone samples.
Ideal for STR Amplification or Massively Parallel Sequencing Workflows
Due to the advantages that massively parallel sequencing (MPS) offers when working with challenging samples, numerous casework laboratories have adopted MPS as a technique when typical STR amplification fails to yield a usable profile. DNA purified with the Bone DNA Extraction Kit can be used in MPS applications. A workflow is outlined below as an example.
Twenty-five-year-old tooth samples from siblings were processed using the Bone DNA Extraction Kit and Maxwell® FSC DNA IQ™ Casework Kit. The extracted DNA was subsequently used in a massively parallel sequencing application as outlined above. Results from the PowerQuant® quantitation assay and library coverage details are shown for duplicates of the two samples. Sequencing results yielded 13 variant calls from each sibling sample that were concordant with each other and were assigned to the H2c haplogroup. Sequencing was performed with the prototype version of the PowerSeq® Whole Mito System.
Protocols
Complete Protocol
Specifications
Catalog Number:
What's in the box?
| Item | Part # | Size | Available Separately |
|---|---|---|---|
Demineralization Buffer |
D678A | 1 × 50ml | |
|
Proteinase K (PK) Solution |
MC500B | 1 × 4ml | View Product |
|
1-Thioglycerol |
A208B | 1 × 900μl | |
|
Lysis Buffer |
A826B | 1 × 150ml | View Product |
SDS
Search for SDSCertificate of Analysis
Use Restrictions
Not For Medical Diagnostic Use.Storage Conditions
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Frequently Used With
Maxwell® FSC Instrument
Benchtop instrument for automated DNA extraction from casework samples.
AS4600
Maxwell® FSC DNA IQ™ Casework Kit
Extract DNA from 1–16 casework samples using the Maxwell® FSC Instrument.
AS1550
PowerQuant® System
5-color, 4-target probe-based qPCR assay.
PQ5002, PQ5008, DS1221
PowerPlex® Fusion System
Co-amplification and fluorescent detection of 24 loci.
DC2402, DC2408
