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LIFE SCIENCE TECHNOLOGIES
670 SCIENCE sciencemag.org/custom-publishing
molecular biology
Produced by the Science/AAAS Custom Publishing Office
IMA
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In May 1983, Kary Mullis, then a scientist at the Cetus
Corporation in Emeryville, California, synthesized some
oligonucleotides, mixed them with a small amount of
template DNA, and added a polymerase enzyme and
a few other reagents. After a series of incubations, the
polymerase had copied the template many times over in a chain
reaction, just as Mullis had hoped.
!"##$�������"�##������������ ��������$���$�����$��� �����
and Cetus was swallowed by a series of other companies, but
PCR persisted. Like the DNA templates it targets, PCR has been
��#$���������#$��������������$���#�� �� $���� #�$���
��$�������� ��#����#� ����� �#�����#���������� �����
vendors sell a wide selection of thermostable polymerases, and
undergraduates worldwide learn the technique in introductory
biology courses.
Now marking its 35th anniversary, PCR has become a ubiquitous
laboratory tool. Nonetheless, researchers, engineers, and physi-
�$����� ����$##���$����������� ��#�$��$��������� $� $���������-
pling of a few of these efforts shows just how far PCRís reach has
grown: from dairies to clinics, and from classrooms to outer space.
The launch sequenceThe primary hardware component of PCR is the thermal
cycler, a machine capable of heating and cooling sample tubes
rapidly, and maintaining them at precise temperatures for
����$����� $�����$������"����� ��������������#������
and competition have improved these machines immensely,
they are still relatively heavy, power-hungry, and expensive. As
�� ��"#���������������$��������$�"������� ���#��"#� �
biology has remained stubbornly inaccessible to educators and
unusable in many remote locations.
ìPCR is one of the most important [research] technologies, and
yet it is one of the most limiting when youíre outside of the lab
because of the size and costî of the equipment, says Zeke Alvarez-
Saavedra, geneticist and cofounder of MiniPCR in Cambridge,
Massachusetts. Frustrated by PCRís persistent immobility, Alvarez-
Saavedra teamed with molecular neurobiologist Sebastian Kraves
in 2013 to build a portable thermal cycler.
Most thermal cyclers control their temperatures using Peltier
junctions, thermoelectric devices that can switch rapidly between
����$�������#$������� �"����#�����#�$� ��"���$���� ��$�����$�����
and the components required to operate them keep PCR ma-
chines heavy and greedy for electricity.
Alvarez-Saavedra and Kraves took a different approach, heating
��������#����$�������$� �#�� ��$��$�������� ��$�$#� ��������$���
defrosters found in cars. For cooling, the team uses a simple fan.
A microcontroller drives the heating, cooling, and incubation
cycles. The simpler design made the machine much smaller and
#$���� ������ $�� ����� ��#����#� ������� "������� ���������
too. ìWhen you make something smaller, it has less parts and
the power supply is smaller ... so that helps reduce the cost,î says
Alvarez-Saavedra.
As they had hoped, the low-cost MiniPCR system immediately
appealed to schools. ìGoing through high school without be-
ing able to get close to biotechnology was kind of a pain point
for us [when we were] becoming scientists, so we wanted to do
away with those barriers,î says Kraves. The company has also
developed a small, simple agarose gel electrophoresis system as
a companion product. A complete kit with the MiniPCR thermal
cycler, gel system, and accessories sells for less than USD 1,000,
putting it well within the budgets of many school systems.
Upcoming features
PCR: Thirty-fve years and counting In the 35 years since its invention, polymerase chain reaction
(PCR) has become a standard technique in laboratory biology,
����������������������������������������������������
������������ ����������������By Alan Dove
Proteomics: Big Data Sharing—June 15 Genomics: Pharmacogenomics—September 28 Animal Models: Better Mouse Models—October 12
LIFE SCIENCE TECHNOLOGIES
67 1SCIENCE sciencemag.org/custom-publishing
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Produced by the Science/AAAS Custom Publishing Office
requires substantial computing power and additional equipment,
making qPCR even more complicated and expensive than
standard PCR.
At Biomeme, cofounders Marc DeJohn, Jesse van Westrienen,
and Max Perelman thought that an unrelated trend might help
bring qPCR out of the lab. ìWe thought, ëhey, everybodyís got a
smartphone in their pocket,í [and] we didnít really see anybody
using them for molecular diagnostics,î says van Westrienen. While
addressing the same engineering problems MiniPCR faced with
'()*(������*�������*'����*��)*(������*(��*�' ����('�)* *���'��*
optical sensors via a smartphone application that can both control
and monitor qPCR reactions, and shelf-stable reagent kits for
sample preparation.
��*����*�)��*�)�����*����*')*��������*(��*�)(*�' ����(*(�'��*
... and one of our focuses from day one has been to develop a
very simple product that doesnít require any expertise or special
lab equipment,î says van Westrienen. The result is a catalog of
kits designed for different applications, each with freeze-dried
reagents and primers premeasured in sample tubes.
Perelman says that by using one of the companyís kits, a
minimally trained experimenter can turn a crude sample into
a set of qPCR reactions ready for the thermal cycler in a few
minutes. The machine itself has a dock compatible with standard
smartphone connectors. Biomemeís web-based data portal can
store the resulting raw data, which researchers can analyze online
or download to their own computers.
At about USD 4,000, the current Biomeme device is
considerably more expensive than MiniPCRís system, but
Perelman argues that it offers additional utility: ìYou can
complete everything in under an hour... just left swipe or right
)�'��*'�*(��*���*���*�������*����*(*)��*(��*����'���('�*'�*
real time.î Perelman adds that the company is now working with
�� ��)�*���*���*�� ������(*�)��)*�)*����*�)*����*)�'��(')()*���*
food companies.
The quick and the deadWhile portable PCR is expanding the techniqueís reach
dramatically, researchers worldwide continue to push its
capabilities forward in countless incremental ways as well. The
problem of distinguishing live from dead bacteria provides a
good example.
���*')*����')'(���*)��)'('��*���*)���'��* �*��(��('��*�����'�*
acid sequences, but simply knowing that a particular DNA or
RNA sequence is present doesnít prove itís associated with a live
organism. Thatís a major problem in the food industry, where
both pasteurized and unpasteurized foods will test positive for
pathogenic bacteria by PCR, even though the bacteria are safely
dead in the former and dangerously alive in the latter. As a result,
food labs have long relied on relatively slow, cumbersome culture
�))��)* �*����'('��*(�)('��
DNA crosslinking reagents that can only penetrate dead
���(��'�*���'���*(��*��)(*��!�*�������*'�*(�')*���� *
Crosslinking prevents the dead bacteriaís DNA from amplifying
'�*(��*)��)�����(*���*����('� *"��*��)(*������('�* *(��)�*
reagents was hard to work with, requiring a darkroom and careful
cold storage. More recently, Takashi Soejima and colleagues at
Morinaga Milk Industry in Kanagawa, Japan, have developed
stable, light-tolerant compounds that similarly target dead
bacteria. The teamís most recent work involved
Though schools have become a major market for MiniPCR,
others have been quick to adopt the platform as well. Kraves says
����*)�'��(')()�*��'���*�������)�*���* �*�����'�)*����*���*��(*
(��*)�)(��*(*�)��* (��*���'��*'(*�������*���* �)(��*(*�*(��'�*
own PCR-based assays on the spot rather than send samples to a
remote lab.
The most surprising call, though, came from engineers
working on the International Space Station. ìWe never really
designed the technology to be space-friendly, so we were
surprised when the space agencies ... told us it was very well
��)'����* �*)����*#'��(�$*)��)*%����) *"��*&'�'���*)�)(��*
is now the core component of the ongoing Genes in Space
competition, where secondary school students propose PCR-
based experiments that are then carried out in microgravity
by astronauts on the space station.
Smartphone PCR&'�'���*��)*(��*��)(*������*(*��(*���*'�(*)�����*��(*
they are not the only one trying to make the technique cheaper
and more portable. Ahram Biosystems in Seoul, South Korea,
sells the Palm PCR miniaturized thermal cycler, which is similar
to the MiniPCR device. Meanwhile, Biomeme in Philadelphia,
Pennsylvania, is taking the portable PCR idea to a new level,
extending it to include real-time PCR.
In real-time or quantitative PCR (qPCR), experimenters use
#���)���(*������)*(*��'(�*(��*�����))* *���*����'���('�*
continuously. By measuring the rate at which new DNA copies
appear, researchers can calculate how many template molecules
must have been in the starting sample. They can also read
results directly from the PCR reaction tubes without having to
run agarose gels. All that monitoring and calculating, however,
Like the DNA templates it targets, PCR has
������� ���������� ����������������
into laboratories worldwide.
cont.>
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672 SCIENCE sciencemag.org/custom-publishing
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Produced by the Science/AAAS Custom Publishing Office
palladium-based reagents that selectively interfered with PCR
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References;��� #��1� 2�ED��.��%�D�������Appl. Environ. Microbiol. 82��<=9>?<=@;��A>;<���A��� ���.��1D����et al���Science 230��;9:>?;9:@��;=B:���9��� 3��� � EE �ED���1�� G��� ���C��)������8��)� E ���Hematology 23�
� ;B;?;B<��A>;B���
Ahram Biosystemsahrambio.com
Biodesixwww.biodesix.com
Biomemebiomeme.com
Bio-Radwww.bio-rad.com
Featured participants
Alan Dove is a science writer and editor based in Massachusetts.
East Carolina Universitywww.ecu.edu
LaCAR MDx Technologies www.lacar-mdx.com
MiniPCRwww.minipcr.com
Morinaga Milk Industrywww.morinagamilk.co.jp/english
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SNP Genotyping System
Integrated DNA Technologiesí rhAmp SNP Genotyping System
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2018
SCIENTIFIC
CONFERENCESPresenting the most significant research on cancer etiology, prevention, diagnosis, and treatment
Learn more and register at
AACR.org/Calendar
Cancer Dormancy and Residual Disease
Conference Cochairs: Julio A. Aguirre-Ghiso,
Ann F. Chambers, Cyrus M. Ghajar,
Christoph A. Klein, and Dorothy A. Sipkins
June 19-22, 2018 | Montreal, QC, Canada
Inaugural AACR International Meeting on
Advances in Malignant Lymphoma: Maximizing the
Basic-Translational Interface for Clinical Application
Scientific Committee Chair: Ari M. Melnick
June 22-26, 2018 | Boston, MA
Sixth JCA-AACR Special Joint Conference on
the Latest Advances in Lung Cancer Research:
FromBasic Science to Therapeutics
Organizing Committee: Hiroyuki Mano,
Seiji Yano, Hiroyoshi Nishikawa, Alice T. Shaw,
Roy S. Herbst, and Charles M. Rudin
July 10-12, 2018 | Kyoto, Japan
Pancreatic Cancer: Advances in
Science and Clinical Care
Conference Cochairs: Ronald M. Evans,
Manuel Hidalgo, Steven D. Leach,
Gloria M. Petersen, and Brian M. Wolpin
September 21-24, 2018 | Boston, MA
Second AACR International Conference on
Translational Cancer Medicine
Conference Cochairs: Carlos L. Arteaga,
Carlos Gil M. Ferreira, and Gabriel A. Rabinovich
September 27-29, 2018 | São Paulo, Brazil
Intestinal Stem Cells and Colon Cancer:
Biology to Therapy
Conference Cochairs: Anil K. Rustgi, Johanna Bendell,
Hans Clevers, Christina Curtis, and Owen Sansom
September 27-30, 2018 | Washington, DC
Metabolism and Cancer
Conference Cochairs: Ralph J. Deberardinis, Tak W. Mak,
Joshua D. Rabinowitz, and M. Celeste Simon
September 28-October 1, 2018 | New York, NY
Fourth CRI-CIMT-EATI-AACR International
Cancer Immunotherapy Conference:
Translating Science into Survival
September 30-October 3, 2018 | New York, NY
EACR-AACR-ISCR Conference: The Cutting
Edge of Contemporary Cancer Research
Conference Cochairs: Richard M. Marais,
Eli Pikarsky, and Robert A. Weinberg
October 9-11, 2018 | Jerusalem, Israel
30th Anniversary AACR Special Conference
Convergence: Systems Biology and
Physical Sciences in Oncology
Conference Cochairs: Phillip A. Sharp
and William C. Hahn
October 14-17, 2018 | Newport, RI
11th AACR Conference on The Science of Cancer
Health Disparities in Racial/Ethnic Minorities
and the Medically Underserved
Conference Cochairs: Laura Fejerman,
Scarlett Lin Gomez, Augusto C. Ochoa,
Brian M. Rivers, and Ivis Sampayo
November 2-5, 2018 | New Orleans, LA
EORTC-NCI-AACR Molecular Targets
and Cancer Therapeutics Symposium
Scientific Committee Cochairs: Charles Swanton,
James L. Gulley, and Antoni Ribas
November 13-16, 2018 | Dublin, Ireland
AACR-KCA Joint Conference on
Precision Medicine in Solid Tumors
Program Committee Cochairs: Tae-You Kim
and Charles L. Sawyers
November 15-17, 2018 | Seoul, South Korea
Tumor Immunology and Immunotherapy
Conference Cochairs: James P. Allison,
Lisa M. Coussens, Ira Mellman, and Drew M. Pardoll
November 27-30, 2018 | Miami Beach, FL