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安全信息

T7699

Sigma-Aldrich

Thunderbolt™ GC10™ Electrocompetent Cells

for generation of cDNA libraries and DNA plasmid production

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About This Item

MDL编号:
MFCD05665590
UNSPSC代码:
12352200

等级

for molecular biology

形式

suspension

运输

dry ice

储存温度

−70°C

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一般描述

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.

应用

Suitable for recovery of high quality plasmid DNA and generation of cDNA libraries from plasmid-based vectors

特点和优势

  • 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

组分

  • Thunderbolt GC10 electrocompetent cells, 5 X 80 μL or 5 X 100 μL (T7074)
  • pUC 19 control DNA (10 ng/μL), 10 μL (D2567)

原理

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.

法律信息

GC10 is a trademark of GeneChoice, Inc.
Thunderbolt is a trademark of GeneChoice, Inc.

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闪点(°F)

Not applicable

闪点(°C)

Not applicable

法规信息

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  1. Which document(s) contains shelf-life or expiration date information for a given product?

    If available for a given product, the recommended re-test date or the expiration date can be found on the Certificate of Analysis.

  2. How do I get lot-specific information or a Certificate of Analysis?

    The lot specific COA document can be found by entering the lot number above under the "Documents" section.

  3. How do I find price and availability?

    There are several ways to find pricing and availability for our products. Once you log onto our website, you will find the price and availability displayed on the product detail page. You can contact any of our Customer Sales and Service offices to receive a quote.  USA customers:  1-800-325-3010 or view local office numbers.

  4. What is the Department of Transportation shipping information for this product?

    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. 

  5. Are home-made competent cells as efficient at transformation as purchased cells?

    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.

  6. Are all competent cells suitable for both plasmid production and protein expression?

    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.

  7. What are important considerations when performing bacterial transformation?

    Things to consider when planning bacterial transformation:DNA impuritiesSource of DNAAmount of DNA usedStorage and handling of the competent cells

  8. Can I store the competent cells in the -20C?

    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.

  9. How should I thaw the competent cells?

    Competent cells should be thawed on ice.  The transformation efficiency is dependent on the proper handling of the cells (maintaining cold temperature until transformation).

  10. Can I re-freeze the vial if I do not use the entire aliquot of competent cells?

    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.

  11. Competent cells were left on ice overnight - can they still be used?

    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.

  12. Can I express a recombinant protein that is toxic to the bacteria?

    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.  

  13. What DNA purity do I need for use in transformation?

    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).

  14. Can I grow more competent cells from the stock I purchased?

    Generally, no, because the future generations of cells lose their competency.

  15. My question is not addressed here, how can I contact Technical Service for assistance?

    Ask a Scientist here.

D Lee Taylor et al.
Molecular ecology resources, 8(4), 742-752 (2008-07-01)
High throughput sequencing methods are widely used in analyses of microbial diversity, but are generally applied to small numbers of samples, which precludes characterization of patterns of microbial diversity across space and time. We have designed a primer-tagging approach that
W Florian Fricke et al.
Journal of bacteriology, 190(20), 6779-6794 (2008-08-19)
The increasing occurrence of multidrug-resistant pathogens of clinical and agricultural importance is a global public health concern. While antimicrobial use in human and veterinary medicine is known to contribute to the dissemination of antimicrobial resistance, the impact of microbial communities
Dana G Mordue et al.
Molecular microbiology, 63(2), 482-496 (2006-12-15)
The apicomplexan parasite Toxoplasma gondii is able to suppress nitric oxide production in activated macrophages. A screen of over 6000 T. gondii insertional mutants identified two clones, which were consistently unable to suppress nitric oxide production from activated macrophages. One
Jeffrey Sabina et al.
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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