PMID- 15490443
OWN - NLM
STAT- MEDLINE
DCOM- 20050418
LR  - 20041027
IS  - 0173-0835 (Print)
IS  - 0173-0835 (Linking)
VI  - 25
IP  - 20
DP  - 2004 Oct
TI  - High-speed DNA sequencing by tube-based capillary electrophoresis.
PG  - 3378-86
AB  - We assessed the feasibility of high-speed DNA sequencing by tube-based capillary 
      electrophoresis (TCE) with electrokinetic sample injections. We developed a 
      water-circulated TCE system to control the capillary temperature precisely. Using 
      this system and a ready-made sieving matrix at 50 degrees C, single-stranded DNA 
      size marker fragments were separated at various pairs of the electric field 
      strength, E, of 128-480 V/cm and the capillary effective length, L, of 100-360 
      mm. Assuming the read length (RL) is the fragment size at which the peak width 
      equals the peak interval per base in obtained electropherograms, we estimated the 
      values of RL (E, L), the RL at the pair (E, L). The points in ELz-space, (E, L, 
      RL(E, L)), form a curved surface expressed by z = RL(E, L). Analyzing the contour 
      lines of this curved surface, we determined the pairs of E and L providing target 
      RLs of 300-500 bases within a minimum time. At a pair optimized for a 500-base RL 
      (330 V/cm, 200 mm), one-color sequencing fragments were successfully separated up 
      to 529 bases within 9.6 min. These results demonstrate that high-speed DNA 
      sequencing comparable with that obtained by microfabricated chip-based capillary 
      electrophoresis (MCE) can be achieved with TCE, which is more suitable in 
      automation than MCE.
FAU - Sakai, Tomoyuki
AU  - Sakai T
AD  - Hitachi Ltd., Central Research Laboratory, Tokyo, Japan. 
      sakai-t@crl.hitachi.co.jp
FAU - Sonehara, Tsuyoshi
AU  - Sonehara T
FAU - Goda, Chifumi
AU  - Goda C
FAU - Kohara, Yoshinobu
AU  - Kohara Y
FAU - Anazawa, Takashi
AU  - Anazawa T
LA  - eng
PT  - Journal Article
PL  - Germany
TA  - Electrophoresis
JT  - Electrophoresis
JID - 8204476
SB  - IM
MH  - Electrophoresis, Capillary/*instrumentation/methods
MH  - Sequence Analysis, DNA/instrumentation/*methods
EDAT- 2004/10/19 09:00
MHDA- 2005/04/19 09:00
CRDT- 2004/10/19 09:00
PHST- 2004/10/19 09:00 [pubmed]
PHST- 2005/04/19 09:00 [medline]
PHST- 2004/10/19 09:00 [entrez]
AID - 10.1002/elps.200406073 [doi]
PST - ppublish
SO  - Electrophoresis. 2004 Oct;25(20):3378-86. doi: 10.1002/elps.200406073.