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文献和实验Polymerase III in vitro Transcription
50 ug/ml BSA 2.5 ul 100 mg/ml BSA H2O to 5 ml total volume 20 ml
Polymerase III in vitro Transcription
5 ug/ml tRNA 1.2 ul tRNA (30 ug/ml) Extract Dialysis Buffer (store frozen ―70 deg): 20 mM HEPES, 7.9 100 mM KCl 5 mM MgCl2 1 mM EDTA 20% glycerol 2 mM DTT Protein dilution buffer (5 ml) store -70 deg
Nested Deletions Using Exonuclease-III and Mung Bean Nuclease
and ethanol ppt. Cut with the second enzyme on the side that deletions are to proceed in around 100ul. Heat inactivate the enzyme when complete. A linearized clone and linearized vector (1ug ea.) provide useful markers for the start and end
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