Células competentes One Shot™ MAX Efficiency™ DH5α-T1R
Células competentes One Shot&trade; MAX Efficiency&trade; DH5&alpha;-T1<sup>R</sup>
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Invitrogen™

Células competentes One Shot™ MAX Efficiency™ DH5α-T1R

Las células competentes One Shot MAX Efficieny DH5α-T1R proceden de la conocida cepa DH5α. DH5α-T1R contiene el genotipo tonA, queMás información
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Número de catálogoCantidad
1229701621 x 50 μl
Número de catálogo 12297016
Precio (MXN)
-
Cantidad:
21 x 50 μl
Las células competentes One Shot MAX Efficieny DH5α-T1R proceden de la conocida cepa DH5α. DH5α-T1R contiene el genotipo tonA, que les confiere resistencia al fago T1 y T5. El bacteriófago T1 se extiende rápidamente y lisa los huéspedes de E. coli, que se suelen emplear para la clonación y la construcción de bibliotecas. Ofrecen una seguridad añadida para proteger clones y bibliotecas de gran valor. Esto es especialmente importante para los centros de secuenciación y genomas, donde la infección de fago T1 sería catastrófica. Además de resistencia a fagos, E. coli DH5α-T1R ofrece las ventajas siguientes:

• Preparaciones de ADN más transparentes y mejores resultados en aplicaciones posteriores debido a la eliminación de la digestión inespecífica por parte de endonucleasa I (endA1)
• Reduce los casos de recombinaciones no deseadas en ADN clonado (recA1)
• Transformación eficaz de ADN no metilado de aplicaciones de PCR (hsdR)
• Detección azul/blanca de clones recombinantes que transportan el fragmento alfa de la β-galactosidasa (lacZΔM15)

Formato práctico y eficaz
E. coli DH5-T1R se suministra en el práctico formato de una sola reacción One Shot. Cada tubo contiene una cantidad suficiente de células para una transformación, sin detrimento de la eficacia ni de los ciclos de congelación y descongelación, además de no desperdiciar dinero en células no utilizadas.
Para uso exclusivo en investigación. No apto para uso en procedimientos diagnósticos.
Especificaciones
Resistencia bacteriana a los antibióticosNo
Tramado azul/blancoSí
Clonación de ADN metiladoNo
Clonación de ADN inestableNo es adecuado para clonar ADN inestable
Contiene el episoma F'Carece de episoma F'
Compatibilidad de alto rendimientoNo compatible con alto rendimiento (manual)
Mejora la calidad de los plásmidosSí
Preparación de ADN no metiladoNo es adecuado para preparar ADN no metilado
Línea de productosOne Shot
Tipo de productoCélula competente
Cantidad21 x 50 μl
Reduce la recombinaciónSí
Condiciones de envíoDry Ice
Resistente al fago T1 (tonA)Sí
Nivel de eficiencia de transformaciónAlta eficacia (> 10^9 ufg⁄µ g)
FormatoTubo
EspecieE. coli
Unit SizeEach
Contenido y almacenamiento
Contiene:
• Células competentes One Shot MAX Efficiency DH5α-T1R: 20 viales, 50 µl cada uno (total de 1 ml)
• ADN pUC19 (10 pg/ul): 1 vial, 50 µl
• Medio S.O.C.: 1 frasco, 6 ml

Almacenar las células competentes a -80 °C. Almacene pUC19 DNA a -20 °C. Almacenar el medio S.O.C. a 4 °C o a temperatura ambiente.

Preguntas frecuentes

How do you recommend that I prepare my DNA for successful electroporation of E. coli?

For best results, DNA used in electroporation must have a very low ionic strength and a high resistance. A high-salt DNA sample may be purified by either ethanol precipitation or dialysis.

The following suggested protocols are for ligation reactions of 20ul. The volumes may be adjusted to suit the amount being prepared.

Purifying DNA by Precipitation: Add 5 to 10 ug of tRNA to a 20ul ligation reaction. Adjust the solution to 2.5 M in ammonium acetate using a 7.5 M ammonium acetate stock solution. Mix well. Add two volumes of 100 % ethanol. Centrifuge at 12,000 x g for 15 min at 4C. Remove the supernatant with a micropipet. Wash the pellet with 60ul of 70% ethanol. Centrifuge at 12,000 x g for 15 min at room temperature. Remove the supernatant with a micropipet. Air dry the pellet. Resuspend the DNA in 0.5X TE buffer [5 mM Tris-HCl, 0.5 mM EDTA (pH 7.5)] to a concentration of 10 ng/ul of DNA. Use 1 ul per transformation of 20 ul of cell suspension.

Purifying DNA by Microdialysis: Float a Millipore filter, type VS 0.025 um, on a pool of 0.5X TE buffer (or 10% glycerol) in a small plastic container. Place 20ul of the DNA solution as a drop on top of the filter. Incubate at room temperature for several hours. Withdraw the DNA drop from the filter and place it in a polypropylene microcentrifuge tube. Use 1ul of this DNA for each electrotransformation reaction.

You offer competent cells in Subcloning Efficiency, Library Efficiency and MAX Efficiency. How do these differ?

There are a few exceptions, but in general the difference is in guaranteed transformation efficiency as follows:

Subcloning Efficiency cells are guaranteed to produce at least 1.0 x 10E6 transformants per µg of transformed pUC19 or pUC18 supercoiled plasmid
Library Efficiency cells are guaranteed to produce at least 1.0 x 10E8 transformants per µg pUC19 or pUC18 DNA
MAX Efficiency cells are guaranteed to produce at least 1.0 x 10E9 transformants per µg pUC19 or pUC18 DNA

Do any Invitrogen competent cells contain DMSO in the freezing medium?

Yes, several of our competent cells products are frozen with DMSO. The presence of DMSO (dimethylsulfoxide) will generally be indicated in the MSDS files if you have a question about a particular product, but here is a list of commonly used products that are known to have DMSO in the freezing buffer:

One Shot OmniMAX 2 T1 Phage Resistant Cells, Cat. No. C8540-03

One Shot INV?F' Chemically Competent Cells, Cat. No. C2020-03 and C2020-06

One Shot MAX Efficiency DH5?-T1 Chemically Competent Cells, Cat. No. 12297-016

MAX Efficiency DH5?-T1 Phage Resistant Cells, Cat. No. 12034-013

MAX Efficiency DH5? Chemically Competent Cells, Cat. No. 18258-012

Library Efficiency DH5? Chemically Competent Cells, Cat. No. 18263-012

MAX Efficiency DH5? F'IQ Cells, Cat. No. 18288-019

MAX Efficiency Stbl2Chemically Competent Cells, Cat. No. 10268-019

When should DMSO, formamide, glycerol and other cosolvents be used in PCR?

Cosolvents may be used when there is a failure of amplification, either because the template contains stable hairpin-loops or the region of amplification is GC-rich. Keep in mind that all of these cosolvents have the effect of lowering enzyme activity, which will decrease amplification yield. For more information see P Landre et al (1995). The use of co-solvents to enhance amplification by the polymerase chain reaction. In: PCR Strategies, edited by MA Innis, DH Gelfand, JJ Sninsky. Academic Press, San Diego, CA, pp. 3-16.

Additionally, when amplifying very long PCR fragments (greater than 5 kb) the use of cosolvents is often recommended to help compensate for the increased melting temperature of these fragments.

Find additional tips, troubleshooting help, and resources within our PCR and cDNA Synthesis Support Center.

Citations & References (1)

Citations & References
Abstract
Haemophilus influenzae type b strain A2 has multiple sialyltransferases involved in lipooligosaccharide sialylation.
Authors: Jones Paul A; Samuels Nicole M; Phillips Nancy J; Munson Robert S Jr; Bozue Joel A; Arseneau Julie A; Nichols Wade A; Zaleski Anthony; Gibson Bradford W; Apicella Michael A;
Journal:J Biol Chem
PubMed ID:11842084
The lipooligosaccharide (LOS) of Haemophilus influenzae contains sialylated glycoforms, and a sialyltransferase, Lic3A, has been previously identified. We report evidence for two additional sialyltransferases, SiaA, and LsgB, that affect N-acetyllactosamine containing glycoforms. Mutations in genes we have designated siaA and lsgB affected only the sialylated glycoforms containing N-acetylhexosamine. A mutation ... More