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GV3101 Agrobacterium Electrocompetent Cells

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Description

GoldBio’s GV3101 Agrobacterium Electrocompetent
Cells are optimized for the highest transformation efficiencies which
is ideal for applications requiring high transformation efficiencies,
such as cDNA or gDNA library construction. The GV3101 strain has a C58
chromosomal background with rifampicin resistance and the Ti plasmid
pmp90 (pTiC58DT-DNA) with gentamicin resistance.
GV3101 is also
resistant to Chloramphenicol and this antibiotic cannot be used for
selection.The GV3101 Ti
plasmid has the T-DNA region sequences deleted and transformation with a
binary vector containing the missing T-region results in a functional
T-DNA binary system that allows for transfer of genetic material into a
host plant S genome. Therefore, this system is often used for
Agrobacterium-mediated transformation of several dicots such as Arabidopsis thaliana, tobacco, potato and soybeans as well as monocots like corn.

Test-drive GoldBio’s competent
cells
with our trial sizes
. Each trial size is available in 3 x 50 μl volumes. Look for the TR mark
in the catalog number for trial size packs.

Table 1: Antibiotic selection for GoldBio’s Agrobacterium strains

Antibiotic Selection

Amp

Carb

Chlor

Gent

Kan

Rif

Spect

Strep

Tet

100
µg/ml

100
µg/ml

30
µg/ml

100
µg/ml

30
µg/ml

50
µg/ml

25
µg/ml

50
µg/ml

50
µg/ml

50
µg/ml

GV3101

S

S

R

PR

R

S

R

PR

R

S

EHA 105

S

S

R

R

R

S

R

R

R

S

LBA 4404

S

S

S

n/a

S

S

R

S

R

S

AGL-1

PS

R

S

n/a

S

S

R

S

R

S

C58C1

S

S

S

n/a

S

S

R

S

R

S

S = Sensitive
R = Resistant
PS = Partial Sensitive (Some growth, but no colonies)
PR = Partial Resistance (Small colonies or some growth in concentrated areas)

Product Specifications

Competent cell type: ElectroCompetent

Species: A. tumefaciens

Strain: GV3101

Format: Tubes

Transformation efficiency: ≥1 x 107 cfu/µg pCAMBIA1391z DNA

Blue/white screening: No

Storage/Handling: This product may be shipped on dry ice. GV3101 Agrobacterium Electrocompetent 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.

Genomic Features

  • ≥1 x 107 cfu/µg efficiency with electroporation.

Reagents Needed for One Reaction

  • GV3101 ElectroCompetent Agrobacterium: 25 µl
  • DNA (pCAMBIA1391z, 100 pg/µl): 1 µl
  • Recovery medium: 1 ml

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 ≥5 x 107 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.

Note: A high-voltage electroporation apparatus capable of generating field strengths of 16 kV/cm is required.

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

P Protocol
O Other
C Certificate of Analysis
S Safety Data Sheet

Additional information

CatalogID

CC-207-A, CC-207-B, CC-207-TR