Introduction
The original transformation protocol dates back to 1970 (Mandel and Higa, J. Mol Biol 1970 , 53; 159). Just preparing the cells took two days. With a protocol two hours long, containing multiple steps, it was a revolution in molecular biology. Does this sound familiar? That’s because it’s still the most popular protocol in use today.
RBC HIT Competent Cells 1 Min competent cells are still currently the fastest transforming cells worldwide since it was launched in 2006. Due to our unique production process cells can be transformed to high efficiency in 1-10 minutes using a simple 1 step patented protocol. RBC provides a wide range of popular strains for all your cloning and expression needs.
Transform your work with RBC HIT Competent Cells.
RBC HIT Competent Cell Efficiency Guarantee
RBC HIT Competent Cells should be stored in a -70°C non frost-free freezer. Competent cells are sensitive to cycles of freezing and thawing and exposure to temperature variations should be minimized. Each RBC HIT package comes with the HIT efficiency guarantee. HIT cells have been electronically temperature monitored to ensure -70°C AT ALL TIMES during batch processing, shipping and storage.
RBC HITCompetent Cell Advantage Just Add Ice !
Compared to traditional Commercial Competent Cells
- Revolutionary 1 minute Transformation
- -70°C Monitoring of competent cell shipments
- True 1 step/1 tube process
- Guaranteed efficiency up to 109 cfu/ug plasmid
- Each batch efficiency checked during production
- Economic pricing for regular and high efficiency cells
- Ideal for bench users to automated cloning projects
- Worldwide patent protection
HIT Competent Cell Selection DH5a, JM109, XL1-Blue, BL-21
- JUMBO : Economical
- VALUE 108 : Convenient and Economical
- HIGH 108 : High Cloning Efficiency
- SUPER 109 : Super Cloning Efficiency
- HIT 96-Well : Cloning Libraries
- JUMBO 80 : More Economical at Price/Prep
Observe growth of transformed colonies.
Protocol Comparison Table
| Competent Cell Types | HIT Competent Cells | Traditional Chemically Competent Cells | Electro Competent Cells |
|---|---|---|---|
| Time | 1 min | 1.5~2.5 hours | 1.5~2.5 hours |
| Protocol steps | One No tube change/SOC |
Multiple Tube change/SOC |
Multiple Tube change/SOC |
| Authentic representation | High | Requires recovery step | Requires recovery step |
| Transformation Efficiency | 107~109 /µg pUC19 | 107~109 /µg UC19 | 1011 /µg pUC19 |
| Stability of refrozen cells | High | Low | Low |
| Stability of vortex | Stable | Unknown | Unknown |
For plasmids > 6 Kb transformation efficiency will neccessarily drop due to the increased MW/plasmid. Please follow the extended protocol outlined below.
Modified Protocol for Large Plasmids:
20-min Ice Bath > 1-min 42°C Water Bath > 20-min Ice Bath
Efficiency Levels
HIT Competent Cells tranformation efficiency reaches 107~109 transformation / µg pUCl9 plasmid DNA.
Minimum Efficiencies Guarantee
HIT VALUE 108 minimum efficiency is 5 x 107 transformants/µg pUC19
HIT HIGH 108 series minimum efficiency is 1 x 108 transformants/µg pUC19
HIT SUPER 109 series minimum efficiency is 5 x 108 transformants/µg pUC19
Applications and Genotype
Genotype Table
| Genotypes | Applications | HIT-DH5a | HIT-Blue | HIT-JM109 | HIT-21 |
|---|---|---|---|---|---|
| F- ( 80d lacZ M15) (lacZYA-argF)U169 hsdR17(r – m +) recA1 endA1 relA1 deoR | hsdR17(rk- mk+), recA1, endA1, gyrA96,thi-1,supE44,relA1, ac[F'proAB acIqZDM15Tn10(Tet )] |
F’traD36proA+proB+ lacIq (lacZ)M15 l (lac-proAB) hsdR17 recA1 endA1 relA1 | E.coli B, F-, dcm, ompT, hsdS(rB-mB-), gal (DE3) |
||
| end A | Prevents plasmid degradation during extraction | Yes | Yes | Yes | No |
| recA | Prevents DNA recombination | Yes | Yes | Yes | |
| hsA | Enhances transformation efficiency of selected PCR DNA strands and cDNA libraries | Yes | Yes | No | Yes |
| deoR | Enhances transformation efficiency of high MW plasmids and cosmids | Yes | No | Yes | No |
| LacZ M15 | Blue-White screening | Yes | Yes | Yes | No |
| Lon | Lon Protease Deficient, Improves Protein Yield | No | No | No | Yes |
| ompT | OmpT Protease Deficient, Improves Protein Yield | No | No | No | Yes |
Application Table
| Applications | HIT-DH5a | HIT-JM109 | HIT-Blue | HIT-21 |
|---|---|---|---|---|
| Large Plasmids > 6Kb* | Ideal | Yes | Yes | Yes |
| Subcloning | Ideal | Ideal | Yes | Yes |
| cDNA library | Yes | - | Yes | Yes |
| Fast Growth | - | Ideal | - | - |
| Single Stranded DNA | - | Ideal | - | - |
| Gene Bank | Yes | Ideal | - | - |
| Mutagenesis | Yes | - | - | - |
| Protein Expression | No | No | No | No |
| Blue/White Screen | Yes | Yes | Ideal | Ideal |
