This document describes the features and characteristics of XL1-Blue MRF' supercompetent cells, and provides information about the related reagents and materials.
The document details the contents of the XL1-Blue MRF' Supercompetent Cells kit, including the specific genotype of the cells which are deficient in restriction systems (McrA-, McrCB-hsdSMR-mrr-, HsdR-) and endonuclease activity. It specifies that the cells must be stored at -80°C immediately upon receipt from dry ice shipping and should not be stored in liquid nitrogen to maintain efficiency.
The guide outlines a step-by-step transformation protocol involving the use of 14-ml BD Falcon polypropylene round-bottom tubes, addition of beta-mercaptoethanol, heat pulsing at 42°C for 45 seconds, and recovery in SOC medium. It also describes conditions for blue-white color screening using pUC18 control DNA.
| Guaranteed Efficiency | ≥1.0 × 10^9 cfu/µg pUC18 DNA |
|---|---|
| Antibiotic Resistance | Tetracycline |
| Heat Pulse Temperature | 42°C |
| Heat Pulse Duration | 45-50 seconds |
Competent cells must be placed immediately at the bottom of a -80°C freezer directly from the dry ice shipping container.
No, do not store the cells in liquid nitrogen as they are sensitive to temperature variations.
It is important that 14-ml BD Falcon polypropylene round-bottom tubes are used, as other tubes may be degraded by beta-mercaptoethanol.