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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
Número MDL:
Código UNSPSC:
12352200
Productos recomendados
grado
Molecular Biology
for molecular biology
Formulario
suspension
Condiciones de envío
dry ice
temp. de almacenamiento
−70°C
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Descripción general
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.
Aplicación
Suitable for recovery of high quality plasmid DNA and generation of cDNA libraries from plasmid-based vectors
Características y beneficios
- 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
Componentes
- 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.
Información legal
GC10 is a trademark of GeneChoice, Inc.
Thunderbolt is a trademark of GeneChoice, Inc.
Producto relacionado
Referencia del producto
Descripción
Precios
Código de clase de almacenamiento
12 - Non Combustible Liquids
Clase de riesgo para el agua (WGK)
WGK 1
Punto de inflamabilidad (°F)
Not applicable
Punto de inflamabilidad (°C)
Not applicable
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What is the Department of Transportation shipping information for this product?
1 answer-
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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?
1 answer-
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?
1 answer-
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?
1 answer-
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 answer-
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?
1 answer-
Generally, no, because the future generations of cells lose their competency.
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Can I store the competent cells in the -20C?
1 answer-
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 answer-
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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