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 | ||||||
711212010-FreeZone 12 Liter -84C Console Freeze Dryer- Labconco
A-104270, 3-Part Life-Size Numbered Human Skull Axis Scientific
911015119999 D2012 Plus High Speed Personal Micro-Centrifuge Scilogex
0091F Assorted Fluorescent Polypropylene Reversible 1.5ml And 2.0ml Microcentrifuge Tube Rack, 96 Places (Pack Of 5) Bio Plas
3970401- 4' Purifier Vertical Clean Bench with UV Light and Protection Panel- Labconco
A-105170 Miniature Painted Human Skeleton Axis Scientific
0094F Fluorescent Orange Polypropylene Reversible 1.5ml And 2.0ml Microcentrifuge Tube Rack, 96 Places (Pack Of 5) Bio Plas
D4074- Quick-DNA Microprep Plus Kit- Zymo Research
28506-QIAquick PCR & Gel Cleanup Kit (100)- Qiagen
0096F WELL MICROCENTRIFUGE TUBE RACK, FLUORESCENT YELLOW, 5/PK Bio Plas
D4075- Quick-DNA Midiprep Plus Kit- Zymo Research
V1002-8 Optional eight channel adapter, for use with 200ul pipette tips Accuris
A-104281 Painted Human Skeleton Axis Scientific
A-104406 Complete Disarticulated Human Skeleton Axis Scientific
28506-QIAquick Spin Columns (100)- Qiagen
V0020-F1 Hydrophobic filter, pk/5 ( 1 each included) Accuris
850071 Environmental Quality SD Card Logger Sper Scientific
Q720 WashLab770 Laboratory Bottle Washer Kimadi
A-106193, Didactic Skull On Cervical Vertebrae With Nerves And Arteries Axis Scientific
0092 Blue Polypropylene Reversible 1.5ml And 2.0ml Microcentrifuge Tube Rack, 96 Places (Pack Of 5) Bio Plas
QIAamp DNA Blood Mini Kit (250) - 51106 - Qiagen
A-105176 Disarticulated Half Child Skeleton Axis Scientific
0093F Fluorescent Green Polypropylene Reversible 1.5ml And 2.0ml Microcentrifuge Tube Rack, 96 Places (Pack Of 5) Bio Plas
WashLab790 Laboratory Bottle Washer Kimadi
0095 Pink Polypropylene Reversible 1.5ml And 2.0ml Microcentrifuge Tube Rack, 96 Places (Pack Of 5) Bio Plas
RE301C-O - Rotary evaporator, Glass "C", Oil BathRE301C-O - Yamato
A-104264, 16-Part Mini Torso Axis Scientific
Q620 WashLab760 Laboratory Bottle Washer Kimadi
A-105168 Miniature Human Skeleton Axis Scientific
V1002-S8 Optional eight channel adapter, fine tip stainless steel Accuris
0097 Lavender Polypropylene Reversible 1.5ml And 2.0ml Microcentrifuge Tube Rack, 96 Places (Pack Of 5) Bio Plas 


