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Cat. No.: MPMSD-20 (for 20T)

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

Cat. No.: MPMSD-100 (for 100T)

Description

Muta-Prem™ Multi-Site Directed Mutagenesis Kit is developed based on the Multi-Fragment Recombination Cloning Kit with the addition of high-fidelity PCR Mix. All kit components have been specially optimized to substantially improve key performance indicators including colony yield and mutation efficiency for multi-site mutagenesis. It can also be applied to single-site mutagenesis experiments.

Similar to the Multi-Fragment Recombination Cloning Kit, mutagenesis primers targeting each mutation site should be designed first. A 20-base complementary sequence is added to the 5' end of PCR primers for adjacent mutated fragments, enabling the termini of neighboring PCR amplicons to carry 20 bp homologous overlap sequences. PCR products of all mutated fragments are mixed at an appropriate ratio. Under the action of recombinase, incubation at 50 °C for 15–30 min enables circularization of all mutated fragments into mutant plasmids. The reaction mixture is subsequently transformed and plated, and 2–3 colonies are picked for sequencing to verify successful mutagenesis.

Our Muta-Prem™ Multi-Site Directed Mutagenesis Kit adopts a proprietary recombinase system together with optimized reaction buffer, delivering drastically elevated experimental success rates of over 95%. When no more than four mutation sites are introduced, purified PCR products are not required for the recombination reaction, greatly simplifying experimental workflows.

Features

• Suitable for multi-site and single-site gene mutagenesis
• Up to >95% mutagenesis success rate
• No PCR purification required for ≤4 mutation sites
• One-step homologous recombination at 50 °C within 15–30 min

Application

Simultaneous introduction of multiple point mutations into target genes

Related:

• Muta-Prem™ Single-Site Directed Mutagenesis Kit
• Muta-Prem™ Chemical-Based Site-Directed Mutagenesis Kit
• Muta-Prem™ Plus Site-Directed Mutagenesis Kit
• Muta-Prem™ Multi-Site Directed Mutagenesis Kit
• Muta-Prem™ Random Mutagenesis Kit
• Muta-Prem™ Random Mutagenesis & Seamless Cloning Kit

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).}}