LBA4404 Agrobacterium Chemically Competent Cells

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Description

GoldBio’s LBA4404 Agrobacterium chemically competent cells allow you to obtain high transformation efficiency in applications such as gDNA or cDNA library construction. Our LBA4404 strain harbors a rifampicin resistance (rif) gene. Furthermore, LBA4404 has an octoprine-type Ti plasmid pAL4404 without self-transport function, containing the vir genes. Our LBA4404 strain can be used in genetic transformation of tomato, tobacco and other plants.

LBA4404 chemically competent cells are free of animal-derived products and grown with animal-free media.

Kit Components

  • Competent Cells
  • 1 x 12 mL Recovery Media
  • 1 x 25 µL Control Plasmid (pCAMBIA1391z Control, 10 ng/µL)


Reagents Needed for One Reaction

  • LBA4404 Chemically Competent Agrobacterium: 50 µL
  • DNA (pCAMBIA1391z Control, 10 ng/µL): 5 µL
  • Recovery medium: 1 mL

Storage/Handling

This product may be shipped on dry ice. LBA4404 Agrobacterium chemically competent cells should be stored at -80°C, pCAMBIA1391z 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.

Antibiotic Selection

Table 1: Antibiotic disc sensitivity for GoldBio’s Agrobacterium strains (using standard BD antibiotic discs)

Antibiotic Selection

Amp

Carb

Chlor

Gent

Kan

Rif

Spect

Strep

Tet

GV3101

I

R

R

PR

R

S

R

S

R

S

EHA105

R

R/S

R

n/a

R/S

S

R

S

R

S

LBA4404

S

S

S

n/a

S

S

R

S

R

S

AGL-1

R

R

R

n/a

R/S

S

R

S

R

S

C58C1

R

R

R

n/a

R/S

S

R

S

R

S

S = Sensitive
R = Resistant
R/S = intermediate zones using standard discs.
I = growth in inhibitory zone with standard disc. “Opaque”, not clear zone of inhibition.

Quality Control

Transformation efficiency is tested by using the pCAMBIA1391z control DNA supplied with the kit and using the protocol given below. Transformation efficiency should be ≥1 x 104 CFU/µg pCAMBIA1391z 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

pCAMBIA Plasmid Vector

LBA4404 Agrobacterium Chemically Competent Cells

View Sizes & Pricing

Catalog Number:
CC-107-5x50
CAS Number:
$199.00

For research use only. Not for food, drug, household, or cosmetic use.
Availability:
In stock
Shipping:
$59 Next Day Air + $20 Dry Ice

    Description

    GoldBio’s LBA4404 Agrobacterium chemically competent cells allow you to obtain high transformation efficiency in applications such as gDNA or cDNA library construction. Our LBA4404 strain harbors a rifampicin resistance (rif) gene. Furthermore, LBA4404 has an octoprine-type Ti plasmid pAL4404 without self-transport function, containing the vir genes. Our LBA4404 strain can be used in genetic transformation of tomato, tobacco and other plants.

    LBA4404 chemically competent cells are free of animal-derived products and grown with animal-free media.

    Kit Components

    • Competent Cells
    • 1 x 12 mL Recovery Media
    • 1 x 25 µL Control Plasmid (pCAMBIA1391z Control, 10 ng/µL)


    Reagents Needed for One Reaction

    • LBA4404 Chemically Competent Agrobacterium: 50 µL
    • DNA (pCAMBIA1391z Control, 10 ng/µL): 5 µL
    • Recovery medium: 1 mL

    Storage/Handling

    This product may be shipped on dry ice. LBA4404 Agrobacterium chemically competent cells should be stored at -80°C, pCAMBIA1391z 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.

    Antibiotic Selection

    Table 1: Antibiotic disc sensitivity for GoldBio’s Agrobacterium strains (using standard BD antibiotic discs)

    Antibiotic Selection

    Amp

    Carb

    Chlor

    Gent

    Kan

    Rif

    Spect

    Strep

    Tet

    GV3101

    I

    R

    R

    PR

    R

    S

    R

    S

    R

    S

    EHA105

    R

    R/S

    R

    n/a

    R/S

    S

    R

    S

    R

    S

    LBA4404

    S

    S

    S

    n/a

    S

    S

    R

    S

    R

    S

    AGL-1

    R

    R

    R

    n/a

    R/S

    S

    R

    S

    R

    S

    C58C1

    R

    R

    R

    n/a

    R/S

    S

    R

    S

    R

    S

    S = Sensitive
    R = Resistant
    R/S = intermediate zones using standard discs.
    I = growth in inhibitory zone with standard disc. “Opaque”, not clear zone of inhibition.

    Quality Control

    Transformation efficiency is tested by using the pCAMBIA1391z control DNA supplied with the kit and using the protocol given below. Transformation efficiency should be ≥1 x 104 CFU/µg pCAMBIA1391z 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

    pCAMBIA Plasmid Vector

    Product Specifications

    Catalog ID: CC-107
    Storage/handling: Store Competent Cells at -80°C.
    Blue/White Screening: No
    Competent Cell Type: Chemically Competent
    Species: A. tumefaciens
    Transformation Efficiency: ≥1 x 104 CFU/µg

    CITATIONS

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