Title of article :
Is pulsed electric field still effective for RNA separation in capillary electrophoresis?
Author/Authors :
Li، نويسنده , , Zhenqing and Dou، نويسنده , , Xiaoming and Ni، نويسنده , , Yi and Chen، نويسنده , , Qinmiao and Cheng، نويسنده , , Shuyi and Yamaguchi، نويسنده , , Yoshinori، نويسنده ,
Issue Information :
روزنامه با شماره پیاپی سال 2012
Pages :
6
From page :
274
To page :
279
Abstract :
Pulsed field capillary electrophoresis (PFCE) is a predominant technique to cope with difficulties in resolving large DNA strands, yet it is still unclear whether pulsed electric field is effective for the separation of higher mass RNA. In this paper we focused on the role of pulsed electric field in large RNA fragments analysis by comparing RNA separation performance in PFCE with that in constant field CE. Separation performance in terms of migration mobility, plate numbers, resolution, and selectivity has been tested for the analysis of RNA from 0.1 to 10.0 kilo nucleotide (knt) under different electrophoretic conditions. Denaturation, important to obtain uniform and identifiable peaks, was accomplished by heating the sample in 4.0 M urea prior to analysis and the presence of 4.0 M urea in the electrophoresis buffer. Results demonstrate that unlike DNA in PFCE, the pulsed electric field mainly affects the separation performance of RNA between 0.4 and 2.0 knt. The migration mobility of long RNA fragments is not a strong function of modulation depth and pulsed frequency. Moreover, the logarithm of RNA mobility is almost inversely proportional to the logarithm of molecule size up to 6.0 knt with correlation coefficient higher than 0.99 in all the polymer concentrations measured here. Resonance frequency of RNA in PFCE was also observed. While these initial experiments show no distinct advantages of using PFCE for RNA separation, they do take further step toward characterizing the migration behavior of RNA under pulsed field conditions.
Keywords :
Capillary electrophoresis , Pulsed field capillary electrophoresis , RNA , DNA , Hydroxyethylcellulose
Journal title :
Journal of Chromatography A
Serial Year :
2012
Journal title :
Journal of Chromatography A
Record number :
1515016
Link To Document :
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