Cold Fusion Cloning Kit with Competent Cells

Fast, easy, and efficient, the phosphatase-free, ligation-free Cold Fusion Cloning Kit will take you to transformation-ready DNA in 20 minutes and a single step
  • Fast—get transformation-ready DNA in as little as twenty minutes
  • Easy—a phosphatase-free and ligation-free system that eliminates the need for specific restriction enzymes
  • Efficient—typically achieve >90% positive clones
  • Versatile—add any insert into any site in any vector using manual or automated workflows

Products

Catalog Number Description Size Price Quantity Add to Cart
MC010B-1 Cold Fusion Cloning Kit with Competent Cells 10 Reactions $260
- +
MC096B-1 Cold Fusion Cloning Kit with Competent Cells - 96-well format 96 Reactions $1925
- +
MC100B-1 Cold Fusion Cloning Kit with Competent Cells 20 Reactions $476
- +
MC101B-1 Cold Fusion Cloning Kit with Competent Cells 50 Reactions $1095
- +

Overview

Overview

Quick and easy ligation-free cloning

Fast, easy, and efficient, SBI’s Cold Fusion Cloning is an excellent choice for any cloning project. Whether you're assembling multiple fragments of DNA or simply adding an insert or gBlock® to a vector, the Cold Fusion Cloning Kit will take you to transformation-ready DNA in a single step—just incubate your DNA fragment(s) with linearized vector for five minutes at room temperature and ten minutes on ice. A typical cloning reaction will deliver a successful cloning rate of more than 90%.

  • Fast—get transformation-ready DNA in as little as twenty minutes
  • Easy—a phosphatase-free and ligation-free system that eliminates the need for specific restriction enzymes
  • Efficient—typically achieve >90% positive clones
  • Versatile—add any insert into any site in any vector using manual or automated workflows

“I have designed and made hundreds and hundreds of clones using the traditional and not so traditional methods since I started doing research. I must say the Cold Fusion kit makes it way too easy to design a vector or new construct—it’s a really good product.”
~Hidevaldo B. Machado, PhD., UCLA

How It Works

How It Works

Streamlining vector construction with Cold Fusion Cloning

Cold Fusion Cloning is a quick and easy way to place an insert—or multiple inserts simultaneously—into a linearized vector. The Cold Fusion Master mix contains a proprietary blend of enzymes that prepares DNA fragments ends for sequence-directed alignment. Homologous DNA ends are efficiently fused together and produce vector clones with great accuracy.

The four-step process is as follows (see workflow illustration below):

  1. Linearize your vector and amplify your insert using primers with at least 15 bp of homology to the ends of your linearized vector.
  2. Mix your PCR-amplified inserts with the linearized vector.
  3. Incubate for 5 minutes at room temperature and then 10 minutes on ice to fuse the insert to the linearized vector.
  4. Transform competent cells with the fusion mix. A typical Cold Fusion Cloning reaction delivers 90% positive clones.

How to use SBI’s easy and efficient Cold Fusion Cloning Kit

Supporting Data

Supporting Data

See the results of a better way to clone

Cold Fusion Cloning can clone 3

Figure 1. Cold Fusion Cloning can clone 3′, 5′, and blunt-end fragments efficiently.

Simultaneously insert multiple fragments into a vector in a defined order

Figure 2. Simultaneously insert multiple fragments into a vector in a defined order. Using Cold Fusion technology, the team at SBI built a 4-in-1 construct to generate iPSCs in only three days.

FAQs

Resources

Citations

  • Scott, TG, et al. (2024) TRPS1 modulates chromatin accessibility to regulate estrogen receptor alpha (ER) binding and ER target gene expression in luminal breast cancer cells. PLoS genetics. 2024; 20(2):e1011159. PM ID: 38377146
  • Li, S, et al. (2024) Epithelioid hemangioendothelioma (EHE) with WWTR1::TFE3 gene fusion, a novel fusion variant. Genes, chromosomes & cancer. 2024; 63(2):e23226. PM ID: 38380774
  • Seavey, CN, et al. (2023) Loss of CDKN2A cooperates with WWTR1(TAZ)-CAMTA1 gene fusion to promote tumor progression in epithelioid hemangioendothelioma. Clinical cancer research : an official journal of the American Association for Cancer Research. 2023;. PM ID: 36598859
  • Alexanian, M, et al. (2023) Chromatin Remodeling Drives Immune-Fibroblast Crosstalk in Heart Failure Pathogenesis. bioRxiv : the preprint server for biology. 2023;. PM ID: 36711864
  • Adhikari, P, et al. (2023) Exocytic machineries differentially control mediator release from allergen-triggered RBL-2H3 cells. Inflammation research : official journal of the European Histamine Research Society ... [et al.]. 2023;. PM ID: 36725743
  • Eid, R. (2023) Functional Characterization of HoDI, a DEAD-box RNA Helicase in Toxoplasma gondii, using RNA-Protein Tethering Assay. Thesis. 2023;. Link: Thesis
  • Rojo-Romanos, T, et al. (2023) Precise excision of HTLV-1 provirus with a designer-recombinase. Molecular therapy : the journal of the American Society of Gene Therapy. 2023;. PM ID: 36934299
  • Jelicic, M, et al. (2023) Discovery and characterization of novel Cre-type tyrosine site-specific recombinases for advanced genome engineering. Nucleic acids research. 2023;. PM ID: 37158248
  • Scott, TG, et al. (2023) TRPS1 modulates chromatin accessibility to regulate estrogen receptor (ER) binding and ER target gene expression in luminal breast cancer cells. bioRxiv : the preprint server for biology. 2023;. PM ID: 37461612
  • Lee, YH, et al. (2023) Sources of Calcium at Connexin 36 Gap Junctions in the Retina. eNeuro. 2023; 10(8). PM ID: 37527925
  • Drumond-Bock, AL, et al. (2023) Increased expression of BRD4 isoforms long (BRD4-L) and short (BRD4-S) promotes chemotherapy resistance in high-grade serous ovarian carcinoma. Genes & cancer. 2023; 14:56-76. PM ID: 37705995
  • Schmitt, LT, et al. (2023) Quantification of evolved DNA-editing enzymes at scale with DEQSeq. Genome biology. 2023; 24(1):254. PM ID: 37932818
  • Koelbel, C, et al. (2023) Development of tandem antigen capture ELISAs measuring QSOX1 isoforms in plasma and serum. Free radical biology & medicine. 2023; 210:212-220. PM ID: 38036070
  • Khanal, S, et al. (2023) A tailed mirtron promotes longevity in Drosophila. Nucleic acids research. 2023;. PM ID: 38048325
  • Yang, YHC, et al. (2022) Innervation modulates the functional connectivity between pancreatic endocrine cells. eLife. 2022; 11. PM ID: 35373736
  • Chen, Z, et al. (2022) ADAMTS9-AS2 regulates PPP1R12B by adsorbing miR-196b-5p and affects cell cycle-related signaling pathways inhibiting the malignant process of esophageal cancer. Cell cycle (Georgetown, Tex.). 2022;:1-16. PM ID: 35503407
  • Thievessen, I, et al. (2022) The focal adhesion protein β-parvin controls cardiomyocyte shape and sarcomere assembly in response to mechanical load. Current biology : CB. 2022;. PM ID: 35688156
  • Riu, M, et al. (2022) Elicitation of Innate Immunity by a Bacterial Volatile 2-Nonanone at Levels below Detection Limit in Tomato Rhizosphere. Molecules and cells. 2022; 45(7):502-511. PM ID: 35791736
  • Srivastava, S. (2022) Domain-specific roles of Gpr126 in ventricular chamber development. Thesis. 2022;. Link: Thesis
  • Wang, R, et al. (2022) Salvianolic acid B suppresses hepatic stellate cell activation and liver fibrosis by inhibiting the NF-κB signaling pathway via miR-6499-3p/LncRNA-ROR. Phytomedicine : international journal of phytotherapy and phytopharmacology. 2022; 107:154435. PM ID: 36155216

Products

Catalog Number Description Size Price Quantity Add to Cart
MC010B-1 Cold Fusion Cloning Kit with Competent Cells 10 Reactions $260
- +
MC096B-1 Cold Fusion Cloning Kit with Competent Cells - 96-well format 96 Reactions $1925
- +
MC100B-1 Cold Fusion Cloning Kit with Competent Cells 20 Reactions $476
- +
MC101B-1 Cold Fusion Cloning Kit with Competent Cells 50 Reactions $1095
- +

Overview

Overview

Quick and easy ligation-free cloning

Fast, easy, and efficient, SBI’s Cold Fusion Cloning is an excellent choice for any cloning project. Whether you're assembling multiple fragments of DNA or simply adding an insert or gBlock® to a vector, the Cold Fusion Cloning Kit will take you to transformation-ready DNA in a single step—just incubate your DNA fragment(s) with linearized vector for five minutes at room temperature and ten minutes on ice. A typical cloning reaction will deliver a successful cloning rate of more than 90%.

  • Fast—get transformation-ready DNA in as little as twenty minutes
  • Easy—a phosphatase-free and ligation-free system that eliminates the need for specific restriction enzymes
  • Efficient—typically achieve >90% positive clones
  • Versatile—add any insert into any site in any vector using manual or automated workflows

“I have designed and made hundreds and hundreds of clones using the traditional and not so traditional methods since I started doing research. I must say the Cold Fusion kit makes it way too easy to design a vector or new construct—it’s a really good product.”
~Hidevaldo B. Machado, PhD., UCLA

How It Works

How It Works

Streamlining vector construction with Cold Fusion Cloning

Cold Fusion Cloning is a quick and easy way to place an insert—or multiple inserts simultaneously—into a linearized vector. The Cold Fusion Master mix contains a proprietary blend of enzymes that prepares DNA fragments ends for sequence-directed alignment. Homologous DNA ends are efficiently fused together and produce vector clones with great accuracy.

The four-step process is as follows (see workflow illustration below):

  1. Linearize your vector and amplify your insert using primers with at least 15 bp of homology to the ends of your linearized vector.
  2. Mix your PCR-amplified inserts with the linearized vector.
  3. Incubate for 5 minutes at room temperature and then 10 minutes on ice to fuse the insert to the linearized vector.
  4. Transform competent cells with the fusion mix. A typical Cold Fusion Cloning reaction delivers 90% positive clones.

How to use SBI’s easy and efficient Cold Fusion Cloning Kit

Supporting Data

Supporting Data

See the results of a better way to clone

Cold Fusion Cloning can clone 3

Figure 1. Cold Fusion Cloning can clone 3′, 5′, and blunt-end fragments efficiently.

Simultaneously insert multiple fragments into a vector in a defined order

Figure 2. Simultaneously insert multiple fragments into a vector in a defined order. Using Cold Fusion technology, the team at SBI built a 4-in-1 construct to generate iPSCs in only three days.

FAQs

Citations

  • Scott, TG, et al. (2024) TRPS1 modulates chromatin accessibility to regulate estrogen receptor alpha (ER) binding and ER target gene expression in luminal breast cancer cells. PLoS genetics. 2024; 20(2):e1011159. PM ID: 38377146
  • Li, S, et al. (2024) Epithelioid hemangioendothelioma (EHE) with WWTR1::TFE3 gene fusion, a novel fusion variant. Genes, chromosomes & cancer. 2024; 63(2):e23226. PM ID: 38380774
  • Seavey, CN, et al. (2023) Loss of CDKN2A cooperates with WWTR1(TAZ)-CAMTA1 gene fusion to promote tumor progression in epithelioid hemangioendothelioma. Clinical cancer research : an official journal of the American Association for Cancer Research. 2023;. PM ID: 36598859
  • Alexanian, M, et al. (2023) Chromatin Remodeling Drives Immune-Fibroblast Crosstalk in Heart Failure Pathogenesis. bioRxiv : the preprint server for biology. 2023;. PM ID: 36711864
  • Adhikari, P, et al. (2023) Exocytic machineries differentially control mediator release from allergen-triggered RBL-2H3 cells. Inflammation research : official journal of the European Histamine Research Society ... [et al.]. 2023;. PM ID: 36725743
  • Eid, R. (2023) Functional Characterization of HoDI, a DEAD-box RNA Helicase in Toxoplasma gondii, using RNA-Protein Tethering Assay. Thesis. 2023;. Link: Thesis
  • Rojo-Romanos, T, et al. (2023) Precise excision of HTLV-1 provirus with a designer-recombinase. Molecular therapy : the journal of the American Society of Gene Therapy. 2023;. PM ID: 36934299
  • Jelicic, M, et al. (2023) Discovery and characterization of novel Cre-type tyrosine site-specific recombinases for advanced genome engineering. Nucleic acids research. 2023;. PM ID: 37158248
  • Scott, TG, et al. (2023) TRPS1 modulates chromatin accessibility to regulate estrogen receptor (ER) binding and ER target gene expression in luminal breast cancer cells. bioRxiv : the preprint server for biology. 2023;. PM ID: 37461612
  • Lee, YH, et al. (2023) Sources of Calcium at Connexin 36 Gap Junctions in the Retina. eNeuro. 2023; 10(8). PM ID: 37527925
  • Drumond-Bock, AL, et al. (2023) Increased expression of BRD4 isoforms long (BRD4-L) and short (BRD4-S) promotes chemotherapy resistance in high-grade serous ovarian carcinoma. Genes & cancer. 2023; 14:56-76. PM ID: 37705995
  • Schmitt, LT, et al. (2023) Quantification of evolved DNA-editing enzymes at scale with DEQSeq. Genome biology. 2023; 24(1):254. PM ID: 37932818
  • Koelbel, C, et al. (2023) Development of tandem antigen capture ELISAs measuring QSOX1 isoforms in plasma and serum. Free radical biology & medicine. 2023; 210:212-220. PM ID: 38036070
  • Khanal, S, et al. (2023) A tailed mirtron promotes longevity in Drosophila. Nucleic acids research. 2023;. PM ID: 38048325
  • Yang, YHC, et al. (2022) Innervation modulates the functional connectivity between pancreatic endocrine cells. eLife. 2022; 11. PM ID: 35373736
  • Chen, Z, et al. (2022) ADAMTS9-AS2 regulates PPP1R12B by adsorbing miR-196b-5p and affects cell cycle-related signaling pathways inhibiting the malignant process of esophageal cancer. Cell cycle (Georgetown, Tex.). 2022;:1-16. PM ID: 35503407
  • Thievessen, I, et al. (2022) The focal adhesion protein β-parvin controls cardiomyocyte shape and sarcomere assembly in response to mechanical load. Current biology : CB. 2022;. PM ID: 35688156
  • Riu, M, et al. (2022) Elicitation of Innate Immunity by a Bacterial Volatile 2-Nonanone at Levels below Detection Limit in Tomato Rhizosphere. Molecules and cells. 2022; 45(7):502-511. PM ID: 35791736
  • Srivastava, S. (2022) Domain-specific roles of Gpr126 in ventricular chamber development. Thesis. 2022;. Link: Thesis
  • Wang, R, et al. (2022) Salvianolic acid B suppresses hepatic stellate cell activation and liver fibrosis by inhibiting the NF-κB signaling pathway via miR-6499-3p/LncRNA-ROR. Phytomedicine : international journal of phytotherapy and phytopharmacology. 2022; 107:154435. PM ID: 36155216