ur guide
For Rearch U Only. Not intended for any animal or human therapeutic or diagnostic u.
pcDNA ™3.1/His A, B, and C
广东师范Catalog number V385-20
Revision date 2 March 2012
Publication Part number 28-0111
MAN0000627
Contents
Kit Contents and Storage (iv)
Methods (1)
Cloning into pcDNA™3.1/His A, B, and C (1)
Transformation and Transfection (6)
年终双薪Appendix (9)
Map of pcDNA™3.1/His A, B, and C Vectors (9)
Features of pcDNA™3.1/His A, B, and C Vectors (10)
Map of pcDNA™3.1/His/lac Z (11)
说明方法及其作用Accessory Products (12)
Technical Support (13)
Purchar Notification (14)
References (15)
Kit Contents and Storage
Shipping and Storage pcDNA™ 3.1/His vectors are shipped on wet ice. Upon receipt, store vectors at –20ºC.
Kit Contents 10 μg each of pcDNA™3.1/His A, B, and C are supplied at 0.5 μg/µL in
10 mM Tris-HCl, 1 mM EDTA, pH 8.0 in a total volume of 20 µL.
10 μg of pcDNA™3.1/His/lac Z is supplied at 0.5 μg/µL in 10 mM Tris-HCl,
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1 mM EDTA, pH 8.0 in a total volume of 20 µL.
For rearch u only. Not intended for any human or animal therapeutic or
diagnostic u.
Methods Cloning into pcDNA™3.1/His A, B, and C
Description of the System pcDNA™3.1/His A, B, and C are 5.5 kb vectors derived from pcDNA™3.1 and
designed for high-level expression and purification of recombinant proteins in
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mammalian hosts. The vectors are supplied in three reading frames to facilitate in frame cloning with a polyhistidine metal-binding tag. The human
cytomegalovirus (CMV) immediate-early promoter provides high-level方差怎么求
expression in a wide range of mammalian cells. In addition, the vector replicates
episomally in cell lines that are latently infected with SV40 or that express the
对外贸易总额SV40 large T antigen (e.g. COS7). High-level stable and non-replicative transient
expression can be carried out in most mammalian cells. The control plasmid, pcDNA™3.1/His/lac Z, is the pcDNA™3.1/His B vector with a 3.2 kb fragment containing the β-galactosida gene cloned in frame with the N-terminal peptide. pcDNA™3.1/His/lac Z is included for u as a positive control for transfection, expression, and purification in the cell line of choice.
General Molecular Biology Techniques For help with DNA ligations, E. coli transformations, restriction enzyme analysis, purification of single-stranded DNA, DNA quencing, and DNA biochemistry, e Molecular Cloning: A Laboratory Manual (Sambrook et al., 1989) or Current Protocols in Molecular Biology (Ausubel et al., 1994).
Maintaining pcDNA™3.1/His Many E. coli strains are suitable for the growth of this vector. To propagate and maintain pcDNA™3.1/His A,B, and C, u the supplied stock solution to transform a r
ec A (recombination deficient), end A (endonuclea A deficient)
E. coli strain like TOP10, TOP10F’, DH5α™-T1R, DH10B™, or equivalent (e page
12 for ordering information). Select the transformants on LB plates containing
50–100 μg/mL ampicillin.
For long-term storage, prepare a glycerol stock of your plasmid-containing E. coli strain.
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