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  • Product Name: Trans1-T1 Phage Chemically Comptent Cell
  • Catalog Number: CD501
  • Storage: at -70 °C for six months
  • 币种:  
    • Specification: 5×100 µl 10×100 µl 20×100 µl
  • Quantity:  

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Overview

Description

Trans1-T1 Phage Resistant Chemically Competent Cell is specifically designed for chemical transformation of DNA. It permits a transformation efficiency of over 109 cfu/μg DNA (tested by pUC19 plasmid DNA).

 

Genotype

F-φ80(lacZ)ΔM15ΔlacX74hsdR(rk-, mk+)ΔrecA1398endA1tonA

 

Features

• High transformation efficiency: >109 cfu/μg (pUC19 DNA).

• Fast-growing, colonies are visible in 8~9 hours.

• Resistance to T1 and T5 phage.

• Blue/white selection.

 

Procedures

•Equilibrate a water bath to 42oC.

•Warm a vial of SOC medium or LB medium to room temperature. Warm selective plates at 37oC for 30 minutes.

•Thaw a vial of 100 μl of Trans1-T1 Phage Resistant Chemically Competent Cell on ice, aliquot 50 μl of the cells into a prechilled 1.5 ml tube, add target DNA (1 to 5 μl ) into the tube. Do not mix by pipetting up and down. Incubate the cells on ice for 30 minutes.

•Heat-shock the cells for 30 seconds at 42oC without shaking. Immediately transfer the tube to ice. Incubate on ice for 2 minutes without shaking.

•Add 500 μl of prewarmed SOC medium or LB medium (without antibiotic) into the tube, mix well and shake at 37oC for 1 hour at 200 rpm for cell recovery and for the expression of antibiotic resistance.

•Spread 20 to 200 μl from each transformation vial on a prewarmed selective plate. The remaining can be stored at 4oC and plated the next day if needed.

•Invert the plate and incubate at 37oC overnight.

•Select colonies and analyze by restriction enzyme digestion, PCR, or sequencing.

 

Notes

•Higher efficiency transformation can be achieved by transforming cells immediately following thawing.

•Avoid repeated thawing.

•Gentle handling is required for the entire procedure.

 

Contents& storage

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Citations & references
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Microsatellite markers for the relictual dove tree, Davidia involucrata (Cornaceae) American Journal of Botany 2.586 Tao C, et al. 2012 Mar http://www.ncbi.nlm.nih.gov/pubmed/22358040
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