Lactococcus lactis IL1403 Electrocompetent Cells
Lactococcus lactis IL1403 Electrocompetent Cells are customer strains optimized for the highest transformation efficiencies which is ideal for applications requiring high transformation efficiencies, such as with cDNA or gDNA library construction.
Competent cell type: ElectroCompetent
Species: Lactococcus lactis
Transformation efficiency: ≥1 x 106 cfu/µg pNZ8148 DNA
Blue/white screening: No
Storage/Handling: This product may be shipped on dry ice. L. lactis IL1403 Electrocompetent Cells should be stored at -80°C, pNZ8148 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.
- ≥1 x 106 cfu/µg efficiency with electroporation.
Reagents Needed for One Reaction
- ElectroCompetent Lactococcus lactis Cells: 25 µl
- DNA (or pNZ8148, 50 ng/µl): 1 µl
- Recovery medium: 1 ml
Transformation efficiency is tested by using the pNZ8148 control DNA supplied with the kit and using the protocol given below. Transformation efficiency should be ≥1 x 106 CFU/µg pNZ8148 DNA. Untransformed cells are tested for appropriate antibiotic sensitivity.
- 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
Transform 1 µl of (50 ng/µl) pNZ8148 control plasmid into 25 µl of cells, add 975 µl of Recovery Medium. Dilute 100 µl of this in 900 µl of Recovery Medium and plate 50 µl. Count the colonies on the plate in two days. If you count 300 colonies, the TE is calculated as follows:
Colonies = 300
µg of DNA = 0.05
Dilution = 50/1000 x 100/1000 = 0.005
TE = 300/0.05/0.005 = 1.2 × 106
Procedure for transformation through electroporation utilizing Lactococcus lactis IL1403 Electrocompetent Cells.
Guide to GoldBio's competent cell collection containing transformation efficiencies, characteristics and applications for each of our competent cell lines.
Chart containing genotypes and transformation efficiencies for GoldBio's competent cell collection.