RR1 Chemically Competent E. coli Cells
GoldBio’s RR1 chemically competent E. coli cells are suitable for high-efficiency transformation in a wide variety of applications such as cloning and sub-cloning. E. coli RR1 is a recA+ derivative of the HB101 strain and can be more useful than HB101 if a recA+ background is required.
Competent cell type: Chemically competent
Derivative of: RR1
Species: E. coli
Transformation efficiency: ≥3 x 107 cfu/µg pUC19 DNA
Blue/white screening: No
Storage/Handling: This product may be shipped on dry ice. RR1 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.
F- Lambda- araC14 leuB6(Am) DE(gpt-proA)62 lacY1 glnX44(AS) galK2(Oc) recA+ rpsL20(strR) xylA5 mtl-1 thiE1 hsdS20(rB-, mB-)
Reagents Needed for One Reaction
- RR1 chemically competent cells: 50 µl
- DNA (or pUC19 Control, 10 pg/µl): 1 µl
- Recovery medium: 1 ml
Transformation efficiency is tested by using the pUC19 control DNA supplied with the kit and using the protocol given below. Transformation efficiency should be ≥3 x 10 7 CFU/µg pUC19 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.
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
|Storage/Handling||Store Competent Cells at -80°C.|