Cat #: 11-727
Zymo Research T3021 Mix & Go! XJa Autolysis Competent Cells, 1ml 500x L-Arabinose, 10 x 100µl/Unit
Cat #: 11-727
Zymo Research T3021 Mix & Go! XJa Autolysis Competent Cells, 1ml 500x L-Arabinose, 10 x 100µl/Unit
1ml 500x L-Arabinose
10 x 100µl/Unit
Brand: Zymo Research- Fast: 80 - 90% of E.coli are lysed in only 10 minutes after harvesting
- Simple 20 Second Transformation: No heat shock! Just add DNA and sprea
- High Transformation Efficiencies: Achieve 108 - 109 transformants perµg of plasmid DNA
$324.50
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1ml 500x L-Arabinose
10 x 100µl/Unit
Brand: Zymo Research- Fast: 80 - 90% of E.coli are lysed in only 10 minutes after harvesting
- Simple 20 Second Transformation: No heat shock! Just add DNA and sprea
- High Transformation Efficiencies: Achieve 108 - 109 transformants perµg of plasmid DNA
While there are many cell lysis methods available to scientists, unfortunately none of these methods combine all the ideal features for simple, efficient, economical, and gentle lysis of E. coli cells. The E. coli XJ autolysing strains from Zymo Research were engineered to address this problem. Mild expression of a chromosomally encoded bacteriophage lambda R gene, encoding the lambda lysozyme, also known as lambda endolysin, is induced during growth. Cells are harvested intact while the peptidoglycan layer of the cell walls has been protected from digestion by the cytoplasmic membrane. The membrane is, however, amenable to disruption by a brief physico-chemical stress such as a freeze-thaw cycle after harvesting the cells. The XJ Autolysis™ method is highly efficient and takes only minutes (unlike traditional multiple freeze-thaw cycles). It can be applied to any number of samples without increasing processing time and labor (unlike sonication or French-press), is reliable and repeatable (unlike lysozyme treatment), and finally, is fully compatible with a wide range of buffers. Additionally, it does not require use of any potentially interfering components such as detergents, commonly found in various lytic buffers. They are also applicable for nucleic acid purification, and available with a DE3 lysogen encoding the T7 polymerase for expressing recombinant proteins driven by the T7 promoter.
| Brand | Zymo Research Corporation |
|---|---|
| Autolysis | Lyses easily. The parent strain JM109 itself will release about 20% of cellular protein after one freeze-thaw cycle. This strain will lyse in a wide range of buffer conditions. 80-90% of E. coli are lysed after a single freeze-thaw treatment. |
| Cell Growth | Grows well, especially when medium is supplemented with 1mM Mg2 . |
| DNA Extraction | This strain is EndA- and yields high quality DNA preparations. |
| DNA Stability | The RecA- mutation in XJa stabilizes repetitive DNA sequences. |
| Processing Time | 10 minutes |
| Transformation Efficiency | 108 - 109 transformants perµg of plasmid DNA |
| Product Storage | -70°C to -80°C |
| Protein Expression | Suitable for general screening, but proteases may degrade small or otherwise unstable recombinant proteins. |
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Q1: Can glycerol be present during the freeze-thaw cycle?
Do not perform the freeze and thaw cycle in a buffer containing glycerol. Glycerol protects the E.coli from forming ice crystals which are essential to the lysis of the cells.
Depending on the amount of material used, the lysed material may become viscous, preventing efficient manipulation. However, for most applications it is not necessary to use a large amount of cell material. If necessary, vortexing vigorously for 30 seconds will decrease viscosity in most cases. Alternatively, a nuclease treatment (e.g. DNAse I) can be used to reduce viscosity. Diluting the cell lysate with additional buffer will also reduce viscosity issues.
If the results obtained are not satisfactory, lysis can be significantly improved by incubating the cells at higher temperatures (25 - 37°C) or for longer time (10 or 20 minutes) after thawing (step 5).
Resuspend the cell pellet in water with or without 0.01% - 0.1% Triton X-100. For His-tag purification, resuspend in the His-Binding Buffer of the His-spin Protein Miniprep kit (Zymo Research product # P2001 or P2002). Acidic buffers and buffers containing higher concentrations of Mg2+ (>1 mM), and related metals that stabilize cell walls, inhibit lysis reaction to a various extent. If possible, add magnesium to the buffer after cells are lysed.
When glucose is added to the growth media, it inhibits the induction of the autolysis genes when it is present in the media. As the cells grow, they consume the glucose as a carbon source. Once the glucose has been consumed autolysis begins.
For best results, cells should not be growing actively prior to arabinose induction. This is achieved by using an overnight starter, where cells are already in the stationary growth phase, as stated in the protocol. If a fresher starter needs to be used, include arabinose already in the starter culture.
All our competent cells are classified into Biosafety level 1 and are not genetic modified organisms. Only when transformed with a plasmid they become GMOs.
Most cloning strains will be dam /dcm unless specifically noted in the genotype.
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