Subcloning of Genes or DNA Fragments

2016 ◽  
pp. 103-126
Keyword(s):  
Dna Fragments

scholarly journals Evaluation of DNA Extraction Methods Developed for Forensic and Ancient DNA Applications Using Bone Samples of Different Age

Genes ◽  
2021 ◽  
Vol 12 (2) ◽  
pp. 146
Author(s):  
Catarina Xavier ◽  
Mayra Eduardoff ◽  
Barbara Bertoglio ◽  
Christina Amory ◽  
Cordula Berger ◽  
...  
Keyword(s):  
Dna Extraction ◽  
Ancient Dna ◽  
Extraction Method ◽  
Fragment Size ◽  
Molecular Genetic ◽  
Extraction Methods ◽  
Degraded Dna ◽  
Size Analysis ◽  
Dna Fragments ◽  
Dna Amount

The efficient extraction of DNA from challenging samples, such as bones, is critical for the success of downstream genotyping analysis in molecular genetic disciplines. Even though the ancient DNA community has developed several protocols targeting small DNA fragments that are typically present in decomposed or old specimens, only recently forensic geneticists have started to adopt those protocols. Here, we compare an ancient DNA extraction protocol (Dabney) with a bone extraction method (Loreille) typically used in forensics. Real-time quantitative PCR and forensically representative typing methods including fragment size analysis and sequencing were used to assess protocol performance. We used four bone samples of different age in replicates to study the effects of both extraction methods. Our results confirm Loreille’s overall increased gain of DNA when enough tissue is available and Dabney’s improved efficiency for retrieving shorter DNA fragments that is beneficial when highly degraded DNA is present. The results suggest that the choice of extraction method needs to be based on available sample, degradation state, and targeted genotyping method. We modified the Dabney protocol by pooling parallel lysates prior to purification to study gain and performance in single tube typing assays and found that up to six parallel lysates lead to an almost linear gain of extracted DNA. These data are promising for further forensic investigations as the adapted Dabney protocol combines increased sensitivity for degraded DNA with necessary total DNA amount for forensic applications.

Human XX males with Y single-copy DNA fragments

Nature ◽  
1984 ◽  
Vol 307 (5947) ◽  
pp. 172-173 ◽  
Author(s):  
Georges Guellaen ◽  
Myriam Casanova ◽  
Colin Bishop ◽  
Danielle Geldwerth ◽  
Gabriel Andre ◽  
...  
Keyword(s):  
Single Copy ◽  
Dna Fragments ◽  
Xx Males ◽  
Copy Dna ◽  
Single Copy Dna

A vector for sequencing long (40-kb) DNA fragments

Gene ◽  
1989 ◽  
Vol 75 (2) ◽  
pp. 315-321 ◽  
Author(s):  
Asad Ahmed
Keyword(s):  
Dna Fragments
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