pcDNA3使用说明.doc

pcDNA3.1(+) pcDNA3.1(-) Catalog nos. V790-20 and V795-20, respectively Version I 081401 28-0104 tech_service@  ii Table of Contents Table of Contents iii Important Information v Purchaser Notification vi Methods 1 Overview1 Cloning into pcDNA3.1 2 Transfection 6 Creation of Stable Cell Lines 7 Appendix 10 pcDNA3.1 Vectors 10 pcDNA3.1/CAT 12 Technical Service13 References15 iii  iv Important Information Contents pcDNA3.1 is supplied as follows: Catalog no. Contents V790-20 20 μg pcDNA3.1(+), lyophilized in TE, pH 8.0 20 μg pcDNA3.1/CAT, lyophilized in TE, pH 8.0 V795-20 20 μg pcDNA3.1(-), lyophilized in TE, pH 8.0 20 μg pcDNA3.1/CAT, lyophilized in TE, pH 8.0 Shipping/Storage Lyophilized plasmids are shipped at room temperature and should be stored at -20°C. Product Qualification Each of the pcDNA3.1 vectors is qualified by restriction enzyme digestion with specific restriction enzymes as listed below. Restriction digests must demonstrate the correct banding pattern when electrophoresed on an agarose gel. The table below lists the restriction enzymes and the expected fragments.  Vector Restriction Enzyme Expected Fragments (bp) pcDNA3.1(+) Nhe I Pst I Sac I 5428 1356, 4072 109, 5319 pcDNA3.1(-) Nhe I Pst I Sac I 5427 1363, 4064 169, 5258 pcDNA3.1/CAT Nhe I Pst I Sac I 6217 2145, 4072 109, 6008 v Purchaser Notification Introduction Use of pcDNA3.1 is covered under a number of different licenses as described below. CMV Promoter Use of the CMV promoter is covered under U.S. Patent Nos. 5,168,062 and 5,385,839 owned and licensed by the University of Iowa Research Foundation and may be used for research purposes only. Commercial users must obtain a license to these patents direc