GB5-alpha™ Chemically Competent E. coli Cells

Request Lot Specific COA

Citation Search Tool

Description

GoldBio’s GB5-alpha™ Chemically Competent E. coli cells are equivalent to DH5-alpha cells and are high efficiency cells suitable for transformation in a wide variety of routine applications such as plasmid isolation, cloning, and subcloning.

GB5-alpha™ chemically competent cells are free of animal-derived products and grown with animal-free media.

GB5-alpha™ Chemically competent E. coli cells are equivalent to DH5-alpha.

GB5-alpha™ Highlights

  • Free of all Animal products
  • High transformation efficiency (≥1 x 108 cfu/µg pUC19 DNA)
  • Ideal for routine cloning and plasmid propagation
  • Widely used in synthetic biology, CRISPR, and mutagenesis workflows
  • Compatible with NEBuilder® HiFi DNA Assembly, Gibson Assembly®, and In-Fusion® Snap Assembly
  • Accepts plasmids and BAC's up to ~15kb
  • Blue-white screening via lacZΔM15 mutation
  • Reduced recombination due to recA1 mutation
  • Low nuclease activity due to endA1 mutation
  • Fast growth and easy handling in the lab

Kit Components

  • Competent Cells
  • 1 x 12 mL Recovery Media
  • 1 x 10 µL Control Plasmid (pUC19 Control, 500 pg/µL)


Reagents Needed for One Reaction

  • GB5-alpha™ chemically competent cells: 50 µL
  • DNA (or pUC19 Control, 500 pg/µL): 2 µL
  • Recovery medium: 1 mL

Storage/Handling

This product may be shipped on dry ice. GB5-alpha™ Chemically Competent E. coli cells should be stored at -80°C, pUC19 Control DNA should be stored at -20°C and recovery medium should be stored at 4°C immediately upon arrival. When stored under the recommended conditions and handled correctly, these products should be stable for at least 1 year from the date of receipt.

Genotype

F– φ80lacZΔM15 Δ(lacZYA-argF)U169 recA1 endA1 hsdR17(rK–, mK+) phoA supE44 λ–thi-1 gyrA96 relA1

Applications

  • Routine plasmid cloning and propagation
  • High-efficiency transformation of recombinant DNA
  • Blue-white screening for identifying successful ligations
  • Construction and screening of DNA libraries (e.g., cDNA, genomic)
  • Cloning guide RNAs (gRNAs) for CRISPR applications
  • Amplification of plasmids after site-directed mutagenesis
  • Propagation of synthetic biology constructs and genetic circuits

Quality Control

Transformation efficiency is tested by using the pUC19 control DNA supplied with the kit and using the protocol given below. Transformation efficiency should be ≥1 x 108 CFU/µg pUC19 DNA. Untransformed cells are tested for appropriate antibiotic sensitivity.

General Guidelines

  • Handle competent cells gently as they are highly sensitive to changes in temperature or mechanical lysis caused by pipetting.
  • Thaw competent cells on ice and transform cells immediately following thawing. After adding DNA, mix by tapping the tube gently. Do not mix cells by pipetting or vortexing.

Calculation of Transformation Efficiency

Transformation Efficiency (TE) is defined as the number of colony forming units (cfu) produced by transforming 1 µg of plasmid into a given volume of competent cells.

  • TE = Colonies/µg/Dilution
    • Colonies = the number of colonies counted
    • µg = amount of DNA transformed in µg
    • Dilution = total dilution of the DNA before plating
Example: Transform 1 µL of (10 pg/µL) control plasmid into 25 µL of cells, add 975 µL of Recovery Medium. Dilute 10 µL of this in 990 µL of Recovery Medium and plate 50 µL. Count the colonies on the plate the next day. If you count 250 colonies, the TE is calculated as follows:

Colonies = 250
µg of DNA = 0.00001
Dilution = 10/1000 x 50/1000 = 0.0005
TE = 250/0.00001/0.0005 = 5.0 × 1010

pUC Plasmid Vector

GB5-alpha™ Chemically Competent E. coli Cells

View Sizes & Pricing

Catalog Number:
CC-101-5x50
CAS Number:
$61.00 $48.00

Availability:
In Stock
Shipping:
Shipping calculated at checkout

    Description

    GoldBio’s GB5-alpha™ Chemically Competent E. coli cells are equivalent to DH5-alpha cells and are high efficiency cells suitable for transformation in a wide variety of routine applications such as plasmid isolation, cloning, and subcloning.

    GB5-alpha™ chemically competent cells are free of animal-derived products and grown with animal-free media.

    GB5-alpha™ Chemically competent E. coli cells are equivalent to DH5-alpha.

    GB5-alpha™ Highlights

    • Free of all Animal products
    • High transformation efficiency (≥1 x 108 cfu/µg pUC19 DNA)
    • Ideal for routine cloning and plasmid propagation
    • Widely used in synthetic biology, CRISPR, and mutagenesis workflows
    • Compatible with NEBuilder® HiFi DNA Assembly, Gibson Assembly®, and In-Fusion® Snap Assembly
    • Accepts plasmids and BAC's up to ~15kb
    • Blue-white screening via lacZΔM15 mutation
    • Reduced recombination due to recA1 mutation
    • Low nuclease activity due to endA1 mutation
    • Fast growth and easy handling in the lab

    Kit Components

    • Competent Cells
    • 1 x 12 mL Recovery Media
    • 1 x 10 µL Control Plasmid (pUC19 Control, 500 pg/µL)


    Reagents Needed for One Reaction

    • GB5-alpha™ chemically competent cells: 50 µL
    • DNA (or pUC19 Control, 500 pg/µL): 2 µL
    • Recovery medium: 1 mL

    Storage/Handling

    This product may be shipped on dry ice. GB5-alpha™ Chemically Competent E. coli cells should be stored at -80°C, pUC19 Control DNA should be stored at -20°C and recovery medium should be stored at 4°C immediately upon arrival. When stored under the recommended conditions and handled correctly, these products should be stable for at least 1 year from the date of receipt.

    Genotype

    F– φ80lacZΔM15 Δ(lacZYA-argF)U169 recA1 endA1 hsdR17(rK–, mK+) phoA supE44 λ–thi-1 gyrA96 relA1

    Applications

    • Routine plasmid cloning and propagation
    • High-efficiency transformation of recombinant DNA
    • Blue-white screening for identifying successful ligations
    • Construction and screening of DNA libraries (e.g., cDNA, genomic)
    • Cloning guide RNAs (gRNAs) for CRISPR applications
    • Amplification of plasmids after site-directed mutagenesis
    • Propagation of synthetic biology constructs and genetic circuits

    Quality Control

    Transformation efficiency is tested by using the pUC19 control DNA supplied with the kit and using the protocol given below. Transformation efficiency should be ≥1 x 108 CFU/µg pUC19 DNA. Untransformed cells are tested for appropriate antibiotic sensitivity.

    General Guidelines

    • Handle competent cells gently as they are highly sensitive to changes in temperature or mechanical lysis caused by pipetting.
    • Thaw competent cells on ice and transform cells immediately following thawing. After adding DNA, mix by tapping the tube gently. Do not mix cells by pipetting or vortexing.

    Calculation of Transformation Efficiency

    Transformation Efficiency (TE) is defined as the number of colony forming units (cfu) produced by transforming 1 µg of plasmid into a given volume of competent cells.

    • TE = Colonies/µg/Dilution
      • Colonies = the number of colonies counted
      • µg = amount of DNA transformed in µg
      • Dilution = total dilution of the DNA before plating
    Example: Transform 1 µL of (10 pg/µL) control plasmid into 25 µL of cells, add 975 µL of Recovery Medium. Dilute 10 µL of this in 990 µL of Recovery Medium and plate 50 µL. Count the colonies on the plate the next day. If you count 250 colonies, the TE is calculated as follows:

    Colonies = 250
    µg of DNA = 0.00001
    Dilution = 10/1000 x 50/1000 = 0.0005
    TE = 250/0.00001/0.0005 = 5.0 × 1010

    pUC Plasmid Vector

    Product Specifications

    Catalog ID: CC-101
    Storage/handling: Store Competent Cells at -80°C.

    CITATIONS

    Featured Articles

    • What Are DH5a Competent Cells?

      What Are DH5a Competent Cells?

      DH5a competent cells are a genetically engineered Escherichia coli (E. coli) strain designed for high-efficiency DNA transformation. DH5a is a go-to strain for DNA cloning, plasmid propagation,...

      Read more

    • Explain Alpha-Complementation and Blue-White Screening To Me Like I’m 10

      Explain Alpha-Complementation and Blue-White Screening To Me Like I’m 10

      Alpha-complementation – it’s a big phrase, but it’s not terribly hard to understand. The technical explanation of alpha-complementation involves combining two parts of a protein,...

      Read more

    • E. coli Cloning Strains for Molecular Biology: What They Are and Why They Matter

      E. coli Cloning Strains for Molecular Biology: What They Are and Why They Matter

      Read more

    • All About Differential Media in Gene Cloning - What it is and How it Works

      All About Differential Media in Gene Cloning - What it is and How it Works

      Differential media is culture media containing compounds that enable visual distinction between the microbes growing on it. These media leverage the distinct metabolic and biochemical...

      Read more

    • Cómo Resolver Problemas De Transformación Bacteriana?

      Cómo Resolver Problemas De Transformación Bacteriana?

      La transformación bacteriana es un procedimiento rutinario en laboratorios de biología molecular. Después de la transformación, solo deberás proceder con la reacción de ligación en...

      Read more

    • Understanding Competent Cells for Bacterial Transformation

      Understanding Competent Cells for Bacterial Transformation

      Transformation is a common method in molecular biology with many applications, including cloning, DNA sequencing, and DNA library construction. To perform transformation, you must use...

      Read more

    Login

    Forgot your password?

    Don't have an account yet?
    Create account