Transcription of Champion pET SUMO Protein Expression System
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User Manual Champion pET SUMO Protein Expression System For high -level Expression and enhanced solubility of recombinant proteins in E. coli and cleavage of native Protein Catalog no. K300-01 Rev. Date: 18 June 2010 Manual part no. 25-0709 MAN0000440 ii iii Table of Contents Kit Contents and Storage ..v Accessory T7-Regulated Expression ..3 Experimental Outline ..5 Methods .. 6 Cloning Considerations ..6 Producing PCR Products ..8 Cloning into pET Transforming One Shot Mach1 -T1R Competent Cells ..10 Analyzing Expressing the PCR Product ..13 Analyzing Samples ..16 Purifying the Recombinant Fusion Using SUMO Troubleshooting ..22 Performing the Control Reactions ..25 Gel Purifying PCR Addition of 3 A-Overhangs Post-Amplification.
2 0.01% gelatin 100 μl dNTP Mix 12.5 mM dATP ... Platinum® Taq DNA Polymerase 500 reactions 10966-034 100 units 10342-053 250 units 10342-012 Taq DNA Polymerase, Recombinant 500 units 10342-020 Platinum® Taq DNA Polymerase High 100 units 11304-011 Fidelity 500 units 11304-029 Kanamycin Sulfate 5 g 11815-024 25 g 11815-032 ...
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