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 λ R gene, encoding the λ lysozyme, also known as λ 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.
| Technical specifications | |||||||
| 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 1 mM Mg2+ | ||||||
| DNA Extraction | This strain is EndA- and yields high quality DNA preparations. | ||||||
| DNA Stability | The RecA- mutation in XJa stabilizes repetitive DNA sequences | ||||||
| Genotype | F`[traD36 proA+B+ laclq ∆(lacZ)M15] ∆(lac-proAB) glnV44 (supE44)e14- (McrA-) thi gyrA96 (NalR) endA1 hsdR17(rK- mK+) relA1 recA1 ΔaraB::λR, cat (CmR), λ(DE3) | ||||||
| Processing Time | 10 minutes | ||||||
| Product Storage | -70°C to -80°C | ||||||
| Protein Expression | Suitable for general screening, but proteases may degrade small or otherwise unstable recombinant proteins. | ||||||
| Transformation Efficiency | 108– 109transformants per µg of plasmid DNA | ||||||
				
RE801C-W- Rotary Evaporator - Yamato							
RE601C-O- Rotary EvaporatorRE601C-O - Yamato							
88870101, Digital and Touchscreen Dry Bath Accessories Thermo Scientific - 46 x 6mm dia. Block							
RE301B-WV - Rotary evaporator, Glass "B", Water Bath, VR300 - Yamato							
SF2200- Stat Fax®2200- Awareness Technology							
A-105172 Human Skeleton With Flexible Spine, Muscle Insertions, And Ligaments Axis Scientific							
R1056- Quick-RNA Midiprep Kit 25 preps- Zymo Research							
A-105176 Disarticulated Half Child Skeleton Axis Scientific							
A-105940, 22 Part Osteopathic Natural Bone Human Skull Axis Scientific							
A-104267 Classic Human Skeleton With Study & Numbering Guide Axis Scientific							
A-105852 Classic Flexible Human Skeleton Axis Scientific							
911015119999 D2012 Plus High Speed Personal Micro-Centrifuge Scilogex							
A-105173 Miniature Skull Axis Scientific							
A-105759, Life-Size Human Skull With Removable Muscles Axis Scientific							
A-105177 Painted And Numbered 3-Part Life-Size Human Skull Axis Scientific							



