In 1970, Mandel and Hige found that Escherichia coli cells can absorb λ bacteriophage DNA when treated with CaCl₂ solution, and this state of cells that can absorb DNA is called the competent state. Competent cells are cells that are induced by physicochemical methods to change the state of the cell membrane, enhance the permeability of the cell membrane, and temporarily change the surface properties of the cell membrane to facilitate the entry of foreign genes or vectors into the recipient cells. Such cells in the optimal state of uptake and accommodation of foreign DNA are called competent cells. Competent cells are essential tools for gene cloning and protein expression. Traditional competence has low transformation efficiency and a long transformation time. Yeasen new competent, more rapid, more efficient.

 

1. What's the difference between fast transformation and regular transformation?
2. What is the efficiency of the fast competent cells?
3. How to choose the appropriate transformation procedure in the experiment?
4. Other cloning-related products of Yeasen

 

1. What's the difference between fast transformation and regular transformation?

Figure 1. Quick conversion (10 min) operation process

Figure 2. Quick heat shock conversion (25 min) operation process

Figure 3. Conventional transformation process

 

2. What is the efficiency of the fast competent cells?

As you can see from the product data, our new fast competent cells have a fast sensitivity and accelerate without reducing efficiency. Yeasen's new fast competent cells help you to stay one step ahead of the cloning process.

Table 1. Products Application Cases

Plasmid	Pfast (add 1ng)	10min	1	>5000
			2	>5000
		25min	1	4061
			2	3862
		Conventional	1	6121
			2	5306
Linked products	6kb	10min	1	1562
			2	1006
		25min	1	940
			2	1120
		Conventional	1	1561
			2	1373

Figure 4. Transformation plate of F-DH5α plasmid

A: pFast plasmid 10 min rapid transformation plate (A-1/A-2 in two replicates); B: pFast plasmid was rapidly transformed into plates for 25 min (B-1/B-2 in two replicates); C: Conventional transformation plates with pFast plasmids (C-1/C-2 in two replicates).

Figure 5. Transformation plate of F-DH5α enzyme-linked products

A: The enzyme-linked products were rapidly transformed into plates for 10 min (A-1/A-2 in two replicates); B: the rapid transformation of the enzyme-linked products for 25 min (B-1/B-2 in two replicates); C: Enzyme-linked products were routinely transformed on plates (C-1/C-2 in two replicates).

 

3. How to choose the appropriate transformation procedure in the experiment?

Fast competence can adapt to both fast and regular procedures, so what procedure should we choose in the actual operation?
In general, fast competent cells can be transformed either rapidly or routinely by heat shock. In the construction of plasmids > 7kb, conventional transformation methods can be used to improve transformation efficiency.
The efficiency of rapid competent cells is higher when coated with ampicillin/carbenicillin-resistant plates, but the efficiency is reduced when coated with kanamycin or other antibiotics because kanamycin is more toxic to the cells without the addition of an incubation step. To improve the transformation efficiency of kanamycin or other resistant plasmids, the incubation step can be increased, that is, the 25 min transformation step or the conventional transformation step can be selected.

 

4. Other cloning-related products of Yeasen

The related products that Yeasen can provide are shown in Table 2.

Table 2.  Relevant recommendation

Name	Product code	Application
TOP10 Chemically Competent Cell TOP10	11801ES	Gene cloning
BL21(DE3) Chemically Competent Cell BL21(DE3)	11804ES	Protein expression
Hieff Clone™ Plus One Step Cloning Kit (inquire)	10911ES	Single fragment rapid cloning directed cloning
Hieff Clone™ Universal One Step Cloning Kit	10922ES	Single/multiple fragment rapid cloning directed cloning
Hieff Clone™ Zero TA Cloning Kit	10907ES	Gene sequencing, replication, and preservation
Hieff Clone™ Zero TA Simple Cloning Kit, MCS-Depleted (inquire)	10908ES	Subcloning
Hieff Clone™ Zero Blunt Cloning Kit	10909ES	Gene sequencing、replication and preservation
Hieff Clone™ Zero Blunt Simple Cloning Kit (inquire)	10910ES	Subcloning
2×Hieff™ PCR Master Mix (With Dye)	10102ES	TA cloning, colony PCR, and genotype identification
2×Hieff™ Ultra-Rapid HotStart PCR Master Mix (with Dye)	10157ES	Various target genes were amplified rapidly

 

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