Can I use LB medium supplemented with ampicin to resuscitate competent cells after transformation?

It should not be added.

Heat shock is to make the bacteria expand instantly, and the surface proteins fall off to form a channel, so that the plasmid can enter, and this process is harmful to bacteria. The bacteria are fragile after 42 degrees of heat shock. In addition, some bacteria will die during heat shock, and the dead bacteria will lyse and release some proteases or something, which is not good for living bacteria.

A LB37 temperature bath without antibiotics is used to allow for better recovery of the bacteria that are still alive. If antibiotics are added, it can increase the burden of bacterial resuscitation. However, the time of the warm bath should not be too long, generally between half an hour and an hour.

Trans 1-T1 Phage Resistant Competent Cells were the fastest growing competent cells at present, and clones were visible at 8-9 h on ampicillin plates. It was used for the screening of blue and white spots, and the blue spots were visible in 12 h; Monoclones cultured overnight were cultured in 2 ml of LB medium for 4-5 h for small plasmid extraction; It is suitable for efficient DNA cloning and plasmid amplification, reducing the occurrence of cloned DNA homologous recombination, and improving the yield and quality of plasmid DNA. T1, T5 phage resistance. Using the PUC19 plasmid, the transformation efficiency was up to 109 cfu g.

No, if the recovery time is short, it will result in concentrations that are lower than those that were not resuscitated, so that colonies cannot grow on the medium.

Summary. Yes, but it requires a specific medium, such as the addition of certain selective antibiotics.

Yes, but it requires a specific medium, such as the addition of certain selective antibiotics.

You've done a great job! Can you elaborate on that?

Competent cells are cells of a special state that require special culture conditions to grow and proliferate. During the transformation of competent cells, a series of changes occur in their morphology and posture impulsive state, which also changes their requirements for nutrients and culture media. If resistant media are used to nurture these transformed competent cells, their growth and proliferation may be affected.

In addition, if the resistant anti-cell substance used has a toxic effect on the transformed cells, it may greatly affect the viability and viability of the cells. Therefore, I recommend using a non-resistant medium to nurture competent cells to ensure normal growth and proliferation.