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 | XJb lysis efficiency is 10-20 % lower than XJa. For optimal lysis, more care needs to be taken when selecting the lysis buffer. However, even very low concentrations of detergent may improve lysis significantly. | ||||||
| Cell Growth | A very robust strain, reaching higher OD’s than E. coli K-strains. | ||||||
| DNA Extraction | XJb is not optimal for DNA extraction. | ||||||
| DNA Stability | This strain is RecA positive. | ||||||
| Genotype | F- ompT hsdSB(rB – mB -) gal dcm ΔaraB::ΛR, cat (CmR) | ||||||
| Processing Time | 10 minutes | ||||||
| Product Storage | -70°C to -80°C | ||||||
| Protein Expression | XJb is ideal for recombinant protein expression. It lacks Lon and OmpT proteases, leading to higher protein yields. | ||||||
| Transformation Efficiency | 108– 109transformants per µg of plasmid DNA | ||||||
MAA228HU22-Anti-AMH Mouse Monoclonal Antibody [clone: 4-1#]- Cloude Clone
FA10005M-PIPETMAN L P200L, 20-200 µL, Metal Ejector-Gilson
81-12120-00, Omega Lum™ W Imaging System Gel Company
3970221- 2' Purifier Vertical Clean Bench with UV Light and Protection Panel- Labconco
4316 Polypropylene 0.5ml Graduated Flat Top Microcentrifuge Tube, Red Bio Plas
A-104274, Functional Knee Joint Axis Scientific
RVX5 - Rat Caging, Innocage® Lids External Water Bottle, Autowatering Innocage® IVC Rat Dual Filter - Innovive
F81045-PIPETMAN M P1200M BT Connected-Gilson
BSW50 Block For Digital Dry Bath - 5 x 50ml Tubes Benchmark Scientific
BCLO3016 Paper Roll For Bluetooth Printer Bas Alliance Bio Expertise
A-104283, 18-Part Premium Unisex Life-Size Torso with Open Back Axis Scientific
F81040-PIPETMAN M P10M BT Connected-Gilson
CNA-TACH-DHH7-Hand Held Tachometer- LW Scientific
OR-C-250-T - Microtube Tray (8 Position) For Use With 250 Ml Cup Tip- Omni International
MAA133RB21-Anti-TNFa Mouse Monoclonal Antibody [clone: D3]-Cloud-Clone
BSWPCR2 Block, 1 x 96-well PCR plate (2&4 block models only) Benchmark Scientific
BSH100-02 Block, 40 x 0.2ml (or 5 x PCR strips) Benchmark Scientific
BSWPCR1 Block, 1 x 96-well PCR plate (Single block model only) Benchmark Scientific
FA10004M-PIPETMAN L P100L, 10-100 µL, Metal Ejector-Gilson
OR-C-250 - 250 ml Cup Tip For Ultrasonic Homogenizer- Omni International
MAA183HU21 -Anti-GLa Mouse Monoclonal Antibody [clone: C8] -Cloud-Clone
PMB 163 -PMB Moisture Analyzer- Adam Equipment
MAA133Hu22-Anti-TNFa Mouse Monoclonal Antibody [clone: C1]- Cloude Clone
911015119999 D2012 Plus High Speed Personal Micro-Centrifuge Scilogex
M283 -Basic Lab Microscope- Unico
LAB 214e -Luna Analytical Balances- Adam Equipment
SPB 723e -Solis Precision Balances- Adam Equipment
QIAamp DNA Blood Mini Kit (250) - 51106 - Qiagen 


