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 | ||||||
A-105939, 22 Part Osteopathic Didactic Human Skull Axis Scientific
QIAamp DNA Blood Mini Kit (250) - 51106 - Qiagen
BCLO3015 Inox Protection Head Alliance Bio Expertise
DC4020- DRYCHILL™ 40 X 1.5/2.0ML- Benchmark
F144075-PIPETMAN G Multichannel P12x300G, 30-300µL-Gilson
GR146-1 -5X TBE Buffer 1LT- Hoefer
BCLO3003 Autoclavable Plastic Stick Grille - 90mm Box Alliance Bio Expertise
711212010-FreeZone 12 Liter -84C Console Freeze Dryer- Labconco
9120010-801 Micrometer reticule Labomed
3970223- 2' Purifier Vertical Clean Bench with Airflow Monitor- Labconco
F81051-PIPETMAN M P12X20M BT Connected-Gilson
BCLO3017 IQ/OQ protocol (French version) Alliance Bio Expertise
700201030- FreeZone 2.5 Liter -50C Benchtop Freeze Dryer- Labconco
F81043-PIPETMAN M P200M BT Connected-Gilson
M153- Buttockmate- Sakamoto
BGWR3005 Baggywel - Fraser Demi - 2 X 5l Alliance Bio Exterpise
S58L Premier Academic Kinesiology Skeleton, Painted And Labeled, Sacral Mount On Mobile Stand- Denoyer Geppert
3970302- 3' Purifier Filtered PCR Enclosure with UV Light and Protection Panel- Labconco
F81045-PIPETMAN M P1200M BT Connected-Gilson
BSH100-500 Block, 2 x 50ml Benchmark Scientific
M280 -Basic Lab Microscope- Unico
BGWR3001 Baggywel - Eau Peptonee Tamponnee - 2 X 5l Alliance Bio Exterpise
BSWCMB Quick-Flip Block Benchmark Scientific
701222030- FreeZone 12 Liter -50C Console Freeze Dryer with Stoppering Tray Dryer- Labconco
BSH100-1213 Block, 15 x 12/13mm diameter tubes & vacutainers Benchmark Scientific
F123789-PIPETMAN Fixed F400, 400 µL-Gilson
F123603-PIPETMAN Classic P5000-Gilson
EPB 2103i -Equinox Precision Balances- Adam Equipment
0094F Fluorescent Orange Polypropylene Reversible 1.5ml And 2.0ml Microcentrifuge Tube Rack, 96 Places (Pack Of 5) Bio Plas
0093 Green Polypropylene Reversible 1.5ml And 2.0ml Microcentrifuge Tube Rack, 96 Places (Pack Of 5) Bio Plas
0096 Well Microcentrifuge Tube Rack, Yellow, 5/Pk
0092F Fluorescent Blue Polypropylene Reversible 1.5ml And 2.0ml Microcentrifuge Tube Rack, 96 Places (Pack Of 5) Bio Plas
0095F Fluorescent Pink Polypropylene Reversible 1.5ml And 2.0ml Microcentrifuge Tube Rack, 96 Places (Pack Of 5) Bio Plas
0091F Assorted Fluorescent Polypropylene Reversible 1.5ml And 2.0ml Microcentrifuge Tube Rack, 96 Places (Pack Of 5) Bio Plas
D4075- Quick-DNA Midiprep Plus Kit- Zymo Research
R1056- Quick-RNA Midiprep Kit 25 preps- Zymo Research
911015119999 D2012 Plus High Speed Personal Micro-Centrifuge Scilogex 



