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文献和实验to alleviate problems with base stacking due to G-C compressions. The following method describes the production of 200ml of a 25% formamide, 6% acrylamide sequencing gel mix but it can be scaled accordingly. You can also vary the formamide concentration upto 40
Inverse PCR & Cycle Sequencing
(Deionized formamide) to 1 part (25 mM EDTA; 50 mg/ml Blue Dextran) 5) Load 1.9 ul onto gel Pharmacia ALF Sequencers have also been used for direct cycle sequencing of PCR products using dye primer technology. Good results were obtained using
acetate (pH 7.0) 1.0 M mercaptoethanol 100 μg/ml tRNA HZ Stop 0.3 M sodium acetate 0.1 M EDTA 25 μg/ml tRNA Loading buffer 80% (v/v) formamide 50 mM Tris-borate (pH 8.3) 1 mM EDTA 0.1% (w/v) xylene cyanol 0.1% (w/v) bromophenol blue.Hints
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