PMID- 16652175
OWN - NLM
STAT- MEDLINE
DCOM- 20070524
LR  - 20071203
IS  - 1473-0197 (Print)
IS  - 1473-0189 (Linking)
VI  - 6
IP  - 5
DP  - 2006 May
TI  - On-chip pressure injection for integration of infrared-mediated DNA amplification 
      with electrophoretic separation.
PG  - 601-10
AB  - Poly(dimethylsiloxane) (PDMS) membrane valves were utilized for diaphragm pumping 
      on a PDMS-glass hybrid microdevice in order to couple infrared-mediated DNA 
      amplification with electrophoretic separation of the products in a single device. 
      Specific amplification products created during non-contact, infrared (IR) 
      mediated polymerase chain reaction (PCR) were injected via chip-based diaphragm 
      pumping into an electrophoretic separation channel. Channel dimensions were 
      designed for injection plug shaping via preferential flow paths, which aided in 
      minimizing the plug widths. Unbiased injection of sample could be achieved in as 
      little as 190 ms, decreasing the time required with electrokinetic injection by 
      two orders of magnitude. Additionally, sample stacking was promoted using laminar 
      or biased-laminar loading to co-inject either water or low ionic strength DNA 
      marker solution along with the PCR-amplified sample. Complete baseline resolution 
      (Res = 2.11) of the 80- and 102-bp fragments of pUC-18 DNA marker solution was 
      achieved, with partially resolved 257- and 267-bp fragments (Res = 0.56), in a 
      separation channel having an effective length of only 3.0 cm. This resolution was 
      deemed adequate for many PCR amplicon separations, with the added advantage of 
      short separation time-typically complete in <120 s. Decreasing the amount of 
      glass surrounding the PCR chamber reduced the DNA amplification time, yielding a 
      further enhancement in analysis speed, with heating and cooling rates as high as 
      13.4 and -6.4 degrees C s(-1), respectively. With the time requirements greatly 
      reduced for each step, it was possible to seamlessly couple IR-mediated 
      amplification, sample injection, and separation/detection of a 278-bp fragment 
      from the invA gene of <1000 starting copies of Salmonella typhimurium DNA in 
      approximately 12 min on a single device, representing the fastest PCR-ME 
      integration achieved to date.
FAU - Easley, Christopher J
AU  - Easley CJ
AD  - Department of Chemistry, University of Virginia, McCormick Road, Charlottesville, 
      VA 22904, USA.
FAU - Karlinsey, James M
AU  - Karlinsey JM
FAU - Landers, James P
AU  - Landers JP
LA  - eng
GR  - R01-HG001832/HG/NHGRI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
DEP - 20060327
PL  - England
TA  - Lab Chip
JT  - Lab on a chip
JID - 101128948
RN  - 0 (DNA, Bacterial)
SB  - IM
MH  - *DNA Replication
MH  - DNA, Bacterial/*genetics/*isolation & purification
MH  - Electrophoresis/*methods
MH  - Infrared Rays
MH  - Microarray Analysis/*methods
MH  - Polymerase Chain Reaction
MH  - Pressure
MH  - Salmonella typhimurium/genetics
EDAT- 2006/05/03 09:00
MHDA- 2007/05/26 09:00
CRDT- 2006/05/03 09:00
PHST- 2006/05/03 09:00 [pubmed]
PHST- 2007/05/26 09:00 [medline]
PHST- 2006/05/03 09:00 [entrez]
AID - 10.1039/b600039h [doi]
PST - ppublish
SO  - Lab Chip. 2006 May;6(5):601-10. doi: 10.1039/b600039h. Epub 2006 Mar 27.