First-dimension gel

MZ Minjie Zhang
KL Kongpan Li
JB Jianhui Bai
WV Willem A. Velema
CY Chengqing Yu
RD Ryan van Damme
WL Wilson H. Lee
MC Maia L. Corpuz
JC Jian-Fu Chen
ZL Zhipeng Lu
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Prepare 8% 1.5 mm thick denatured first-dimension gel using the UreaGel system (National Diagnostics, EC-833). Loading dsRNA ladder (NEB, N0363S) as molecular weight marker. Run the first-dimension gel at 30 W for 7–8 min in 0.5× TBE (Invitrogen, 15581044). After electrophoresis was finished, staining the gel with SYBR Gold (Invitrogen, S11494) in 0.5× TBE and excising each lane between 50 nt to topside from the first-dimension gel. The second-dimension gel can usually accommodate three gel splices.

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