For expression of toxic recombinant proteins, select the BL21 strain containing the pLysS plasmid. These cells express low levels of lysozyme that will bind to T7 polymerase, and inhibit transcription.
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Wichtige Dokumente
T7699
Thunderbolt™ GC10™ Electrocompetent Cells
for generation of cDNA libraries and DNA plasmid production
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About This Item
MDL-Nummer:
UNSPSC-Code:
12352200
Empfohlene Produkte
Qualität
for molecular biology
Form
suspension
Versandbedingung
dry ice
Lagertemp.
−70°C
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Allgemeine Beschreibung
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.
Anwendung
Suitable for recovery of high quality plasmid DNA and generation of cDNA libraries from plasmid-based vectors
Leistungsmerkmale und Vorteile
- 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
Komponenten
- Thunderbolt GC10 electrocompetent cells, 5 X 80 μL or 5 X 100 μL (T7074)
- pUC 19 control DNA (10 ng/μL), 10 μL (D2567)
Prinzip
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.
Rechtliche Hinweise
GC10 is a trademark of GeneChoice, Inc.
Thunderbolt is a trademark of GeneChoice, Inc.
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WGK
WGK 1
Flammpunkt (°F)
Not applicable
Flammpunkt (°C)
Not applicable
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Can I express a recombinant protein that is toxic to the bacteria?
1 answer-
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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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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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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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Are home-made competent cells as efficient at transformation as purchased cells?
1 answer-
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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How should I thaw the competent cells?
1 answer-
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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What is the Department of Transportation shipping information for this product?
1 answer-
Transportation information can be found in Section 14 of the product's (M)SDS.To access the shipping information for this material, use the link on the product detail page for the product.
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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 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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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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