Highlights
- •Compilation of current literature for rapid PCR techniques for human identification.
- •Uses of rapid PCR techniques within integrated rapid DNA testing devices.
- •Examination of rapid PCR on chip platforms with/without capillary electrophoresis.
Abstract
Multiplex PCR with fluorescently labeled primers has been an essential method for
the amplification of short tandem repeats used in human identify testing. Within the
STR workflow of extraction, quantitation, amplification, separation, and detection,
multiplex PCR is commonly identified as the bottleneck in the process. The time requirement
of up to three hours to complete 28–30 cycles of multiplex PCR for STR genotyping
is the greatest amount of time required for a single step within the process. The
historical use of commercially available thermal cyclers and heat stable polymerases
may have given the impression that large multiplex will always require long PCR cycling
times to ensure that all of the varying sized targets (typically 100–400 bp) can be amplified in a balanced manner throughout the multiplex. However, with
the advent of improved polymerases and faster thermal cyclers the time required for
the amplification of large STR multiplexes is no longer on the order of three hours,
but as little as 14 min. Faster amplification times can be performed while retaining the balance and integrity
of large multiplex PCRs by implementation of alternate polymerases and thermal cyclers.
With the reduction in PCR cycling times there has also been an impact on the development
of integrated and microfluidics devices which employ the use of reduced or rapid thermal
cycling protocols as part of their integration. Similarly, PCR inhibitor resistant
polymerases can also reduce overall STR processing times for reference samples by
eliminating the need for DNA extraction and purification that is additionally implemented
within the development of integrated DNA typing devices.
Keywords
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Article info
Publication history
Published online: April 23, 2015
Accepted:
April 21,
2015
Received in revised form:
April 3,
2015
Received:
February 18,
2015
Identification
Copyright
Published by Elsevier Inc.
