XJa (DE3) Autolysis Glycerol Stock
- Fast: 80 - 90% of E.coli are lysed in only 10 minutes after harvesting.
- DE3 Lysogen: Encodes the T7 polymerase for expressing recombinant proteins under the control of the T7 promoter.
- Versatile: Fully compatible with a wide range of buffers for protein purification and other physical methods of lysis.
|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.|
Q1: Is a starter culture necessary?
Q2: What buffer should the cell pellet be resuspended in?
Q3: How do you improve lysis efficiency?
Q4: What if the lysate is extremely viscous?
Q5: Can glycerol be present during the freeze-thaw cycle?
Q6: Can glucose be added to the growth media?
Q7: Will chitin be degraded?
Q8: Are competent cells GMOs?
Q9: Which is the recommended DNA concentration and volume for transformation?
Q10: Which Plasmid Size can be used for transformation?
Q11: Which antibiotics can be used with the Mix & Go! procedure?
Q12: Is it possible to dilute the competent cells?
Q13: How to reduce satellite colonies on agar plates?
Q14: How will a heat-shock affect my Transformation Efficiency?
Q15: Do the Mix & Go! strains methylate DNA?
Q16: Are the Mix & Go! strains dam+ and dcm+?
Q17: What are some tips to improve transformation efficiency?
Q18: Which strains are equivalent to the Zymo strains?
To clone new GFP-like fluorescent proteins from Obelia medusa, the authors identified the potential genes using expression libraries and cloned the genes into a vector. Expression of the proteins was facilitated by using XJb Autolysis E. coli cells from Zymo Research. The authors were able to purify three proteins from Obelia medusa that fluoresce in three different colors: cyan, green, and yellow.Aglyamova, G.V. et al. (2011) Multi-colored homologs of the green fluorescent protein from hydromedusa Obelia sp. Photochem Photobiol Sci (8):1303-9.