Scale-up development of high-performance polymer matrix for DNA sequencing analysis.
Linear polyacrylamide (LPA) has been widely used as a replaceable separation matrix in CE. An increase in the molecular weight of the separation medium favors the separation of larger DNA fragments. In order to obtain ultrahigh-molecular-weight (UHMW) LPA, a "frozen" method was developed to synthesize the LPA homopolymer. This approach has three major advantages when compared with other existing routes of LPA synthesis: (i) long LPA chains could be obtained easily, with their average molecular weight (MW) being in the high 10 MDa range; (ii) the desired MW could be adjusted over a broad range by controlling the temperature and the concentration of initiators during synthesis; (iii) the product solution contains only a tiny amount of impurity besides the solvent and LPA. Both static and dynamic laser light scattering measurements were carried out to characterize the synthesized LPA in the buffer solution. The DNA sequencing matrix prepared from LPA using this method was studied and the results were compared with the newly developed commercial product POP7 from Applied Biosystems. It should be noted that this approach can be applied to synthesize other water-soluble polymers, resulting in UHMW products because the chain transfer constant is smaller at lower temperatures.
Wan
Fen
F
Department of Chemistry, Stony Brook University, Stony Brook, NY, USA.
He
Weidong
W
Zhang
Jun
J
Ying
Qicong
Q
Chu
Benjamin
B
D000180:Acrylic Resins / Q000138:chemical synthesis* / Q000737:chemistry; D019075:Electrophoresis, Capillary; D005615:Freezing; D008970:Molecular Weight; D011108:Polymers / Q000138:chemical synthesis / Q000737:chemistry; D017422:Sequence Analysis, DNA / Q000379:methods*; D000099166:Polyacrylamides
D000180:Acrylic Resins; D011108:Polymers; D000099166:Polyacrylamides