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Cat. No.: MPRM-50 (for 50T)

Cat. No.: MPRM-200 (for 200T)

Description

Muta-Prem™ Random Mutagenesis Kit is a reagent kit based on Error Prone PCR (epPCR), which introduces a certain probability of random mutations into the target gene fragment through PCR amplification.

Random mutagenesis of genes is an important method for studying the relationship between protein structure and function, as well as inhibiting, modifying, or even creating protein or enzyme activities. The epPCR method typically involves using an error-prone thermostable DNA polymerase for PCR amplification, thereby introducing random mutations into the target gene sequence. After introducing random mutations into the target gene sequence, it is highly likely to result in amino acid sequence mutations at the protein level, leading to proteins or enzymes with enhanced, diminished, or even lost activity. It can also generate new activities in proteins or enzymes.

For analyzing the relationship between protein structure and function, the ideal mutation would be one amino acid change per protein (1-2 nucleotide mutations). In the study of protein evolution, the ideal mutation would be 1-4 amino acid changes per protein (approximately 2-7 nucleotide mutations). When screening highly active proteins or enzymes from a mutation library, it is typically desired to have around 20 nucleotide mutations per gene.

The RND-Muta DNA Polymerase provided in this kit is a novel DNA polymerase specifically designed for error-prone PCR. When the recommended amount of Mutation enhancer is added to the supplied reaction buffer, it can introduce approximately 7.8 random mutations per kilobase of the target gene. These mutations are balanced between AT→GC and GC→AT directions, with a ratio of (AT→GC mutations) / (GC→AT mutations) = 1.06. It also introduces a suitable number of insertion and deletion mutations.

The amplified DNA fragments produced by this kit have sticky ends and can be directly used for conventional T-vector cloning.

This kit can amplify target fragments up to 6 kb in length.

Users have the flexibility to adjust the mutation rate of this kit according to their needs, making it more convenient to use. The kit provides Mutation enhancer (10X), and the mutation rate can be significantly influenced by altering the amount of Mutation enhancer used.

Precautions

• PCR reactions are highly sensitive. When using RND-Muta DNA Polymerase for PCR amplification, be careful to avoid contamination of the template DNA with minute amounts, and it is recommended to include a no-template control (NTC) to account for background amplification.
• This product is intended for scientific research purposes by professionals only. It is not intended for clinical diagnosis or treatment, and should not be used for food or drugs. It should not be stored in regular residential areas.
• For your safety and health, please wear appropriate laboratory attire and disposable gloves when handling this product.

Storage

The minimum shelf life is 1 year at -20°C.

Instruction: Protocol

Featured Citations

Interested in seeing published research using our Mutagenesis Kits?

NLRP6 potentiates PI3K/AKT signalling by promoting autophagic degradation of p85α to drive tumorigenesis

^{_{Nature Communications    |    28 September 2023    |    DOI: https://doi.org/10.1038/s41467-023-41739-z}}

^{_{p85α mutants were generated using a site-directed mutagenesis kit (SBS Genetech, Beijing, China) according to the manufacturer’s instructions.}}

ZmBSK1 positively regulates BR-induced H2O2 production via NADPH oxidase and functions in oxidative stress tolerance in maize

^{_{Plant Physiology and Biochemistry    |      15 August 2022    |    Doi: https://doi.org/10.1016/j.plaphy.2022.06.011}}

^{_{The mutated ZmCCaMK was obtained using the Site-Directed Mutagenesis Kit (SBS Genetech) followed by the manufacturer's protocol.}}

Rhophilin rho GTPase binding protein 1-antisense RNA 1 (RHPN1-AS1) promotes ovarian carcinogenesis by sponging microRNA-485-5p and releasing DNA topoisomerase II alpha (TOP2A)

^{_{Bioengineered    |      07 Dec 2021    |    Doi: https://doi.org/10.1080/21655979.2021.2002494}}

^{_{The site-directed mutagenesis kit (SBS Genetech, China) was used to mutate the WT binding sequence of RHPN1-AS1 or TOP2A 3ʹ-UTR, and the produced mutant sequence was also inserted into psiCHECK2 vectors, which were named RHPN1-AS1 Mut1, RHPN1-AS1 Mut2, RHPN1-AS1 co-Mut, and TOP2A Mut vectors.}}

Thr420 and Ser454 of ZmCCaMK play a crucial role in brassinosteroid-induced antioxidant defense in maize

^{_{Biochemical and Biophysical Research Communications Supports open access    |      7 May 2020    |    Doi: https://doi.org/10.1016/j.bbrc.2020.02.078}}

^{_{The Site-Directed Mutagenesis Kit (SBS Genetech, China) was used to obtain the site-directed ZmCCaMK according to the manufacturer’s instructions.}}

Long non-coding RNA LOC554202 promotes acquired gefitinib resistance in non-small cell lung cancer through upregulating miR-31 expression

^{_{Journal of Cancer    |     15 Oct 2019    |    Doi: https://doi.org/10.7150%2Fjca.35097}}

^{_{The mutation of miR-31 binding sites in 3'-UTRs of RASA1 (UCUUGCC was mutated to UGUUCGG) or FIH-1 (UCUUGCC was mutated to UGUUCGG) was performed by a Muta Direct Site-directed Mutagenesis kit (SDM-15; Beijing SBS Genetech Co, Ltd, Beijing, China).}}

Long Non-coding RNA H19 Inhibits Adipocyte Differentiation of Bone Marrow Mesenchymal Stem Cells through Epigenetic Modulation of Histone Deacetylases

^{_{Scientific Reports    |     28 June 2016    |    DOI: https://doi.org/10.1038/srep28897}}

^{_{Site-directed mutagenesis of the H19 sequences was performed using the Site-Directed Mutagenesis Kit (SBS Genetech, Beijing, China).}}

仅用于研究，不用于治疗人类或动物

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