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) | ||||||
| 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 | ||||||
4303 Polypropylene 1.5ml Graduated Flat Top Microcentrifuge Tube, Blue Bio Plas
0052 Snap On Tube Rack Cover, 96-Well, Polypropylene, Clear Natural Bio Plas
911015119999 D2012 Plus High Speed Personal Micro-Centrifuge Scilogex
A-106193, Didactic Skull On Cervical Vertebrae With Nerves And Arteries Axis Scientific
RE301A-WV - Rotary evaporator, Glass "A", Water Bath, VR300- Yamato
A-104267 Classic Human Skeleton With Study & Numbering Guide Axis Scientific
A-105170 Miniature Painted Human Skeleton Axis Scientific
0093 Green Polypropylene Reversible 1.5ml And 2.0ml Microcentrifuge Tube Rack, 96 Places (Pack Of 5) Bio Plas
4307 Polypropylene 1.5ml Graduated Flat Top Microcentrifuge Tube, Lavender Bio Plas
0099 Black Polypropylene Reversible 1.5ml And 2.0ml Microcentrifuge Tube Rack, 96 Places (Pack Of 5) Bio Plas
RE601A-O - Rotary EvaporatorRE601A-O - Yamato
0091 Polypropylene Reversible 1.5ml And 2.0ml Microcentrifuge Tube Rack, 96 Places (Pack Of 5) Bio Plas
A-105867, Human Skull Model With Flexible Neck Axis Scientific
0094 Orange Polypropylene Reversible 1.5ml And 2.0ml Microcentrifuge Tube Rack, 96 Places (Pack Of 5) Bio Plas
RC 900 - Rotary Evaporator- Logos BioSystem
A-105168 Miniature Human Skeleton Axis Scientific
711212110-FreeZone 12 Liter -84C Console Freeze Dryer- Labconco
A-105172 Human Skeleton With Flexible Spine, Muscle Insertions, And Ligaments Axis Scientific
A-105868, Human Skull With Masticatory Muscles Axis Scientific
88870102, Digital and Touchscreen Dry Bath Accessories Thermo Scientific-28 x 10mm dia. Block
V1002-8 Optional eight channel adapter, for use with 200ul pipette tips Accuris
28506-QIAquick PCR & Gel Cleanup Kit (100)- Qiagen
PTM 500 Drum and Wheelchair Platforms Adam Equipment
A-104264, 16-Part Mini Torso Axis Scientific
RE301A-O - Rotary evaporator, Glass "A", Oil Bath- Yamato
4306 Polypropylene 1.5ml Graduated Flat Top Microcentrifuge Tube, Orange Bio Plas
711222110-FreeZone 12 Liter -84C Console Freeze Dryer with Stoppering Tray Dryer- Labconco
A-105939, 22 Part Osteopathic Didactic Human Skull Axis Scientific 



