Plasmid DNA high-frequency transformation of Escherichia coli experiments
Plasmid DNA high-frequency transformation of Escherichia coli experiments
After the preparation of the receptor cells, the next step is the transformation of plasmid DNA into E. coli cells. However, it is important to note that the receptor cells should preferably be freshly prepared, because the preservation of receptor cells for a certain period of time will reduce the transformation rate; in addition, the concentration of DNA should also be careful, not too high.
Operation method
high frequency method
Materials and Instruments
Receptor Cells Move 1, take a tube of newly prepared sensory cells. For more product details, please visit Aladdin Scientific website.
Plasmid DNA Antibiotics
Ep tubes Water baths LB plates
2, take 0.03 ml of sensory cells and 4 ng of plasmid DNA, mix well, placed in an ice bath for 30 min.
3、Place the Ep tube in a water bath at 42 ℃ for 2 minutes, and then immediately place it on ice for 1 minute.
4、Add 70 ul LB medium to the Ep tube and mix well, incubate at 37 ℃ for 30min.
5、Coat on LB plate containing appropriate concentration of antibiotics.
6、It was incubated at 37 ℃ overnight, and the growth of plaque was regarded as positive clone.