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 | ||||||
710201000-FreeZone 2.5 Liter -84C Benchtop Freeze Dryer- Labconco
D4074- Quick-DNA Microprep Plus Kit- Zymo Research
D4075- Quick-DNA Midiprep Plus Kit- Zymo Research
3970305- 3' Purifier Filtered PCR Enclosure with UV Light, Protection Panel and Airflow Monitor- Labconco
R1056- Quick-RNA Midiprep Kit 25 preps- Zymo Research
QIAamp DNA Blood Mini Kit (250) - 51106 - Qiagen
911015119999 D2012 Plus High Speed Personal Micro-Centrifuge Scilogex
0095 Pink Polypropylene Reversible 1.5ml And 2.0ml Microcentrifuge Tube Rack, 96 Places (Pack Of 5) Bio Plas
28506-QIAquick PCR & Gel Cleanup Kit (100)- Qiagen
0091 Polypropylene Reversible 1.5ml And 2.0ml Microcentrifuge Tube Rack, 96 Places (Pack Of 5) Bio Plas
Q620 WashLab760 Laboratory Bottle Washer Kimadi
V1002-8 Optional eight channel adapter, for use with 200ul pipette tips Accuris
V0020-F1 Hydrophobic filter, pk/5 ( 1 each included) Accuris
711212010-FreeZone 12 Liter -84C Console Freeze Dryer- Labconco
A-106193, Didactic Skull On Cervical Vertebrae With Nerves And Arteries Axis Scientific
700611230- FreeZone 6 Liter -50C Console Freeze Dryer- Labconco
0090 Natural Polypropylene Reversible 1.5ml And 2.0ml Microcentrifuge Tube Rack, 96 Places (Pack Of 5) Bio Plas
3970402- 4' Purifier Filtered PCR Enclosure with UV Light and Protection Panel- Labconco
3970202- 2' Purifier Filtered PCR Enclosure with UV Light and Protection Panel- Labconco
3970205- 2' Purifier Filtered PCR Enclosure with UV Light, Protection Panel and Airflow Monitor- Labconco 


