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T7699
Thunderbolt™ GC10™ Electrocompetent Cells
for generation of cDNA libraries and DNA plasmid production
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About This Item
Numero MDL:
Codice UNSPSC:
12352200
Prodotti consigliati
Grado
for molecular biology
Stato
suspension
Condizioni di spedizione
dry ice
Temperatura di conservazione
−70°C
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Descrizione generale
Thunderbolt GC10 electrocompetent E. coli have a very high transformation efficiency of >1x1010 cfu/μg when transformed with non-saturating amounts of pUC19 plasmid DNA. They are comparable to DH10β strain.
Applicazioni
Suitable for recovery of high quality plasmid DNA and generation of cDNA libraries from plasmid-based vectors
Caratteristiche e vantaggi
- Ensures recovery of stable, high quality plasmid DNA as well as methylated DNA
- Renders protection to clonal stocks from T1 and T5 bacteriophage contamination
- Allows for β-galactosidase α-complementation for blue/white screening
- Guaranteed high transformation efficiency
- Convenient 80 μL or 100 μL aliquots
Componenti
- Thunderbolt GC10 electrocompetent cells, 5 X 80 μL or 5 X 100 μL (T7074)
- pUC 19 control DNA (10 ng/μL), 10 μL (D2567)
Principio
Thunderbolt GC10 electrocompetent E. coli is K strain bacteria that contain mutations in recA1 and endA1 genes. These mutations aid in minimizing recombination and ensuring plasmid stability. This strain also contains tonA genotype that confers resistance to lytic bacteriophages such as T1 and T5. The host restriction systems are eliminated to allow the cloning of methylated DNA.
Note legali
GC10 is a trademark of GeneChoice, Inc.
Thunderbolt is a trademark of GeneChoice, Inc.
Prodotti correlati
N° Catalogo
Descrizione
Determinazione del prezzo
Codice della classe di stoccaggio
12 - Non Combustible Liquids
Classe di pericolosità dell'acqua (WGK)
WGK 1
Punto d’infiammabilità (°F)
Not applicable
Punto d’infiammabilità (°C)
Not applicable
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D Lee Taylor et al.
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W Florian Fricke et al.
Journal of bacteriology, 190(20), 6779-6794 (2008-08-19)
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The Journal of biological chemistry, 284(43), 29635-29643 (2009-09-02)
Efficient uptake of glucose is especially critical to Saccharomyces cerevisiae because its preference to ferment this carbon source demands high flux through glycolysis. Glucose induces expression of HXT genes encoding hexose transporters through a signal generated by the Snf3 and
Plasmids of Escherichia coli as cloning vectors.
F Bolivar et al.
Methods in enzymology, 68, 245-267 (1979-01-01)
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What is the Department of Transportation shipping information for this product?
1 risposta-
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Are home-made competent cells as efficient at transformation as purchased cells?
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They can be, depending on the technique used, the expected transformation efficiency and how the cells are handled. For special applications, competent cells prepared in-house using standard methods may not provide the efficiency you need.
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Can I re-freeze the vial if I do not use the entire aliquot of competent cells?
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Re-freezing competent cells will result in a decrease in their transformation efficiency. If the cells are frozen first in a dry ice / ethanol bath before placement in the -80C freezer, the loss will be about two-fold. If placed directly in the -80C freezer, the loss in transformation efficiency is about five to ten-fold.
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How should I thaw the competent cells?
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Competent cells should be thawed on ice. The transformation efficiency is dependent on the proper handling of the cells (maintaining cold temperature until transformation).
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Are all competent cells suitable for both plasmid production and protein expression?
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No. Most strains of competent cells are suitable for producing plasmid DNA that will be used to transfect insect or mammalian cells for expression of the protein. The BL21 competent cell strains have special traits enabling protein expression in bacteria.
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What are important considerations when performing bacterial transformation?
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Things to consider when planning bacterial transformation:DNA impuritiesSource of DNAAmount of DNA usedStorage and handling of the competent cells
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Competent cells were left on ice overnight - can they still be used?
1 risposta-
Competent cells left on ice, or allowed to come to room temperature slowly will suffer a dramatic loss in transformation efficiency. We recommend thawing a new aliquot of competent cells.
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Can I grow more competent cells from the stock I purchased?
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Generally, no, because the future generations of cells lose their competency.
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Can I store the competent cells in the -20C?
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No. Competent cells should not be stored at -20C for any length of time. The cells suffer a dramatic drop in transformation efficiency when stored higher than -80C.
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What DNA purity do I need for use in transformation?
1 risposta-
The DNA used should be high quality - free from phenol, proteins, detergents, and ethanol. Dissolve the DNA in sterile water or 0.5x TE buffer (5 mM Tris-HCl, 0.5 mM EDTA).
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