Ethanol re-precipitation removes PCR inhibitors from Ancient DNA extract
نویسندگان
چکیده
One of the major problems in ancient DNA work is the presence of inhibitory substances, which hampers Taq polymerase activity. Therefore analysis of ancient DNA sample is very challenging. Here we describe a simple and competent ethanol re-precipitation based protocol for the purification of DNA from ancient bones and tissues. The efficiency of this procedure has been demonstrated on 600 years old biological samples provided by Anthropological Survey of India (Himalaya region). This suggests that re-precipitation of ancient DNA extracts removes PCR inhibitors and increases the success rate of amplification.
منابع مشابه
A simple and efficient method for PCR amplifiable DNA extraction from ancient bones.
A simple and effective modified ethanol precipitation-based protocol is described for the preparation of DNA from ancient human bones. This method is fast and requires neither hazardous chemicals nor special devices. After the powdering and incubating of the bone samples Dextran Blue was added as a carrier for removing the PCR inhibitors with selective ethanol precipitation. This method could e...
متن کاملComparison and optimization of ancient DNA extraction.
Ancient DNA analyses rely on the extraction of the tiny amounts of DNA remaining in samples that are hundreds to tens of thousands of years old. Despite the critical role extraction efficiency plays in this field of research, no study has comprehensively compared ancient DNA extraction techniques to date. There are a wide range of methods currently in use, which rely on such disparate principle...
متن کاملGenomic Dna Purification
The Extract-N-AmpTM Blood PCR Kits contain all of the reagents necessary to rapidly extract host genomic DNA from whole blood and amplify targets of interest by PCR (Fig. 1). This novel extraction system eliminates the need for any type of purification, organic extraction, centrifugation, heating, filtration or alcohol precipitation. The kit also includes a PCR Ready mix, especially formulated ...
متن کاملA simple method for extracting DNA from old skeletal material.
Extraction of DNA from old skeletal material is of great importance in the identification of human remains, but is particularly difficult because the methods currently employed, especially those using phenol/chloroform, are not always satisfactory. A simple technique based on the removal of non-nucleic acid material by salting out (precipitation) with saturated sodium acetate is described; the ...
متن کاملSpiking of contemporary human template DNA with ancient DNA extracts induces mutations under PCR and generates nonauthentic mitochondrial sequences.
Proof of authenticity is the greatest challenge in palaeogenetic research, and many safeguards have become standard routine in laboratories specialized on ancient DNA research. Here we describe an as-yet unknown source of artifacts that will require special attention in the future. We show that ancient DNA extracts on their own can have an inhibitory and mutagenic effect under PCR. We have spik...
متن کامل