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Current Location:Home > Cloning Services > Site-directed Mutagenesis


Site-directed Mutagenesis


Site-directed mutagenesis is a molecular biology method of altering the nucleotide sequence of the target gene at a specified location using the polymerase chain reaction(PCR), including insertions, deletions, point mutations. Site-directed mutagenesis is a commonly-used method in gene research, which can investigate the character and characterization of the target protein in a short time. WZ Biosciences provides the following site-directed mutagenesis services(single or multiple mutation) for your scientific research. you can select one of the clones from our ORF Clone Collection or send your own clones. As long as you tell us about your mutation needs, we can assist you in completing point mutation services and a series of downstream services, including Molecular Cloning 、Virus Packaging (Adenovirus, Lentivirus and AAV) and Gene Expression Analysis, saving your valuable time.


Cat No.

Type

Specification

Price

Turnaround Time

WZ020001

Single mutation

5ug DNA; 500ul Glycerol Stock

Quote

Inquiry

WZ020002

Multiple mutation

5ug DNA; 500ul Glycerol Stock

Quote

Inquiry

1、Any combination of base changes within 6bp is counted as one mutation site;
2、Additional cloning costs are needed for vector exchanging.


*Interested in above Mutagenesis Services? Please feel free to contact us to “Request a quote”


Related services:Molecular Cloning, Adenovirus Packaging, Lentivirus Packaging, Adeno-associated virus(AAV) Packaging, Stable Cell Lines Generation



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Site-directed mutagenesis is used to introduce specific mutation into the target DNA of known sequence. Depending on the base change, site-directed mutagenesis usually includes three types: base substitution, base deletion, and base insertion. According to whether the amino acid changes after the base changes, gene mutations can also be divided into three types: same sense mutations, missense mutations and nonsense mutations.

In basic research, researchers usually create a mutant gene through changing the DNA sequence to investigate the relationship between gene structure and its function; or generate a mutant protein by changing amnio acid sequence to study the relationship of between protein structure and its function. In this way, under physiological conditions, researchers can clarify the regulation mechanism of gene, the etiology and pathogenesis of disease from the micro level.


A most commonly used method for site-directed mutagenesis: PCR

The flow chart of performing site-directed mutagenesis by PCR: 1. Design mutagenic primers; 2. overlap PCR; 3. Template DNA is eliminated by a methylation-dependent endonuclease (i.e. DpnI); 4. The PCR product that are digested by DpnI are transformed into bacteria; 5. Positive clones selecting and sequencing. The flow chart is below:


维真生物 ‖ PCR法进行定点突变的流程示意图
1. Ready-to-Use mutation templates: WZ Biosciences has two extensive human gene collection, including human ORF collection and human miRNA collection.
2. Intimate Follow-Up Services: WZ Biosciences can provide you with a variety of follow-up services such as subcloning, adenovirus packaging, lentivirus packaging, AAV packaging services and gene expression analysis.
3. Fair and reasonable price.

《Transfection Reagent Instructions》

1. What materials and information do I need to provide for site-directed mutagenesis?

The materials to be provided by you:

If you will send your own vector for mutating or cloning, please send us at least 5 ug DNA or glycerol stock;

The information to be provided by you:

①Gene Name, sequence(pre-mutation sequence) and sequencing results;
②Description of sites/base pairs to mutate;
③Name of desired viral vector backbone or name of customer-provided vector.


2. How do you define one mutation?

One mutation is defined as any base changes within 6bp.


3. If I need to change the destination vector while mutating, how do you charge?

If you need to change the destination vector while mutating, additional cloning cost will be required.

Human ORF Clone Human microRNA Clone Human shRNA Clone
Human gRNA Pool Clone Mouse gRNA Pool Clone ……

1.

Aging (Albany NY). (IF=4.831). Li, et al. (2019). Sirt1-inducible deacetylation of p21 promotes cardiomyocyte proliferation.

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