New block-copolymer thermoassociating matrices for DNA sequencing: effect of molecular structure on rheology and resolution.
A new family of matrices for DNA sequencing by capillary electrophoresis is presented. These matrices combine easy injection with high sieving performances, due to thermal switching between a low and a high viscosity state through a modest increase in temperature (approximately 20 degrees C). They are constructed from a hydrophilic polymer backbone with grafted lower critical solution temperature (LCST) side chains. The comb-like LCST copolymers are characterized in terms of size of the polymer backbone, the size of LCST side chains and the grafting densities. The dependance of rheological behavior and electrophoretic performance of these copolymers is correlated with their microstructure. Without complete optimization, a resolution of order 0.5, corresponding to a very reasonable limit for read length with current base calling softwares, could be achieved for segments around 800 bases differing by 1 base in less than one hour in a commercial ABI 310 apparatus.
Sudor
J
J
Laboratoire de physico-chimie Curie (UMR CNRS 168), Institut Curie, Section de Recherche, Paris, France.
Barbier
V
V
Thirot
S
S
Godfrin
D
D
Hourdet
D
D
Millequant
M
M
Blanchard
J
J
Viovy
J L
JL
D000180:Acrylic Resins / Q000138:chemical synthesis / Q000737:chemistry*; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D019075:Electrophoresis, Capillary / Q000379:methods*; D006358:Hot Temperature; D011108:Polymers / Q000737:chemistry*; D012212:Rheology; D017422:Sequence Analysis, DNA / Q000379:methods*; D012996:Solutions; D014783:Viscosity; D014867:Water
D000180:Acrylic Resins; D011108:Polymers; D012996:Solutions; D014867:Water; C052970:poly-N-isopropylacrylamide