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Improvement in the I-PEP method and its effect on the outcome of low copy number DNA profiling
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Abstract: (16170 Views) |
| Background & Aim: DNA Profiling has become one of the most robust and reliable methods at forensic identification However, an insufficient DNA quantity (less than 100 Pg or 33 copies) found often in forensic evidence samples, is a major hindrance. Amplification of such low copy number DNA samples is attainable with the most efficient whole genome amplification (WGA) method, named improved primer extension preamplification (I-PEP) PCR.
Methods: By initial assessment of existing PEP and I-PEP methods on serially diluted DNA, it was attempted to reach an improved method leading to reliable profiling with the lowest amount of template. This method employs degenerate 15-mer PCR primers followed by specific amplification of DNA with specific primers.
Results: Subsequent to the amplification with the new modified and improved I-PEP, which we term KI-PEP PCR, complete DNA profile was obtained from only 2.5 pg of input DNA. Using this method, a fragment size of 1106 bp was effortlessly amplified with the specific primers.
Conclusions: The Utility of KI-PEP PCR not only increases the low quantity of DNA, but also provides the optimum length appropriate to DNA Typing techniques. |
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| Keywords: Medicine; Forensic Identification; DNA Profiling; WGA; KI-PEP PCR |
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Type of Article: Research Article |
Received: 2011/05/14 | Revised: 2011/05/21 | Accepted: 2020/06/10 | ePublished: 2020/06/10
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