Cambio - Excellence in Molecular Biology

Molecular Cloning Kits

Molecular Cloning Kits: RED/ET Kits

Quick and Easy BAC Modification

The Quick and Easy BAC Modification kit is designed to modify any type of bacterial artificial chromosomes (BACs) within 1-2 weeks by using a kanamycin/neomycin cassette

Gene Bridges

Catalogue No.DescriptionPack SizePriceQty
K001Quick and Easy BAC Modification1 kit £568.00 Quantity Add to Order

Quick and Easy BAC Modification

The Quick and Easy BAC Modification kit is designed to modify any type of bacterial artificial chromosomes (BACs) within 1-2 weeks by using a kanamycin/neomycin cassette

Gene Bridges

Overview of one-step insertion of a selectable marker gene

1. Transformation of BAC host withRed/ET
In a first step the Red/ET plasmid pSC101-BAD-gbaA is transferred into the E.coli host that contains the BAC.
quickandeasybac 1
2. BAC Modification by Red/ET
The expression of genes mediating Red/ET is induced by adding L-arabinose and a temperature shift from 30°C to 37°C. After induction the cells are prepared for electroporation and the PCR product with the added homology arms is electroporated
quickandeasybac 2
3. Selection & Extraction of the modified BAC DNA
Select for the antibiotic resistance conveyed by the selectable marker in the PCR product to identify colonies carrying successfully modified BACs. A subsequent DNA mini preparation is used to confirm the desired BAC modification.
quickandeasybac 3


This kit is designed to modify BACs with the revolutionary Red/ET Recombination technology - a powerful tool for introducing modifications such as insertions and deletions into any type of BAC within 2 weeks.

Red/ET Recombination uses in vivo recombination and proofreading activity to minimize unwanted mutations in your construct. Included in the kit are all the necessary contents for modifying BACs and all materials needed to perform a control experiment intended to familiarize you with this novel technique.

Additionally, a vector converting any E.coli strain into competent strains for application with Red/ET Recombination is provided. pSC101-BAD-gbaAtet comes with a tetracyclin resistance cassette. This cassette contains a temperature sensitive origin of replication (ori) that can only propagate at 30°C - at 37°C the plasmid will be lost. This allows for simple removal of the plasmid when it is no longer needed.

To introduce a mutation (i.e. a selection cassette like the neomycin resistance cassette included with this kit) you first have to introduce homology sequences to the selection cassette. This can be done through a simple PCR reaction.
These short homologous sequences (50 nt each) can be freely chosen for your experimental needs. Simply choose the location to be modified and insert the selection cassette directly at the desired nucleotide with the Red/ET Recombination technology. The PCR product then needs to be electroporated in bacteria with the induced Red/ET Recombination system.

After successful Red/ET Recombination the modified DNA can be recovered using conventional DNA mini-preparation methods.


  • pSC101-BAD-gbaA (tet) which is a derivative of pSC101 ori (a temperature sensitive origin) based Red/ET expression plasmid
  • BAC host carrying pSC101-BAD-gbaA (tet)
  • Positive control experiment for modifying a 150kb BAC (BAC clone in the host carrying pSC101-BAD-gbaA, neo PCR product for deletion in BAC backbone, BAC-neo clone in the host as positive recombinant)
  • Protocols, descriptions of plasmids, maps and sequences of oligos


Tn5-kanR/neoR selection cassette

Tn5-promoter  kanR/neoR



One Year Non-Commercial License Agreement


Purchase orders submitted for kits, plasmids or strains that contain a gene encoding any of RecE, RecT, Reda and Redb can not be processed without acceptance of the "One Year Non-Commercial License Agreement for Red/ET Recombination Technology"

If you cannot find the answer to your problem below then please contact us or telephone 01954 210 200

Quick and Easy BAC Modification

The Quick and Easy BAC Modification kit is designed to modify any type of bacterial artificial chromosomes (BACs) within 1-2 weeks by using a kanamycin/neomycin cassette

Gene Bridges

Protocols for: Quick & Easy BAC Modification Kit

Quick & Easy BAC Modification Kit Protocol

(catalogue number K001) 

Please note: all protocols off site are the responsibility of the products supplier

If you cannot find the answer to your problem below then please contact us or telephone 01954 210 200

Quick and Easy BAC Modification

The Quick and Easy BAC Modification kit is designed to modify any type of bacterial artificial chromosomes (BACs) within 1-2 weeks by using a kanamycin/neomycin cassette

Gene Bridges

Gene Bridges Publications

Ambrogio C, Stern P, Scuoppo C, Kranz H, Barbacid M and Santamaría D, 2014, Lentiviral-based approach for the validation of cancer therapeutic targets in vivo, BioTechniques, 57, 179

Vaisman A, McDonald JP, Noll S, Huston D, Loeb G, Goodmann MF and Woodgate R, 2014, Investigating the mechanisms of ribonucleotide excision repair in Escherichia coli, Mutation Res, 761, 21

Noll S, Reyelt J, Rysiok T, Kellner, R, Güssow D, Jäkel S, Hager S and Kranz H, 2013, Gezielte Optimierung von Escherichia coli BL21(DE3), Biospektrum, 19, 211

Hager S, Lösch S, Noll S, Khan-Vaughan L, Ehrlich ME and Kranz H, 2012, Red/ET recombination with chimeric oligonucleotides allows rapid generation of BAC transgene harboring full-length or truncated huntingtin cDNA, BioTechniques Rapid Dispatches, doi: 10.2144/000113908

Noll S, Hampp G, Bausbacher H, Pellegata NS and Kranz H, 2009, Site-directed mutagenesis of multi-copy-number plasmids: Red/ET recombination and unique restriction site elimination, BioTechniques, 46, 527

Braatsch S, Helmark S, Kranz H, Koebmann B and Jensen PR, 2008, Escherichia coli strains with promoter libraries constructed by Red/ET recombination pave the way for transcriptional fine-tuning, BioTechniques, 45, 335

Fu J, Wenzel SC, Perlova O, Wang J, Gross F, Tang Z, Yin Y, Stewart, AF, Müller R and Zhang Y, 2008, Efficient transfer of two large secondary metabolite pathway gene clusters into heterologous hosts by transposition, Nucleic Acids Research, 36, 17

Heermann R, Zeppenfeld T and Jung K, 2008, Simple generation of site-directed point mutations in the Escherichia coli chromosome using Red/ET Recombination, Microbial Cell Factories 7, 14

Lösch S, Braatsch S and Kranz H, 2007, Red/ET Recombination. λ vs. Mu: base-precise modification of the E. coli genome, BIOspektrum

Wang J, Sarov M, Rientjes J, Fu J, Hollak H, Kranz H, Xie W, Stewart AF and Zhang Y, 2006, An improved recombineering approach by adding RecA to lambda Red recombination, Mol Biotechnol, 32, 43

Zeppenfeld T and Kranz H, 2006, Manipulation des E. coli-Chromosoms zur Optimierung von Produktionsstämmen, Laborwelt, 5, 25

Zhang Y, Stevens G, Stewart AF, 2002, Ein neues Prinzip der DNA-Klonierung, LaborPraxis, 72



2013/2014: Strain modification & metabolic engineering approaches

Satoh et al., 2014, New gene responsible for para-aminobenzoate biosynthesis, J Bioscience and Bioeng, 117, 178

Jung et al., 2013, Removal of pathogenic factors from 2,3-butanediol-producing Klebsiella species by inactivating virulence-related wabG gene, Apppl Gen & Mol Biotech, 97, 1997

Wang et al., 2013, Engineering furfural tolerance in Escherichia coli improves the fermentation of lignocellulosic sugars into renewable chemicals, PNAS, 110, 4021

Pfander et al., 2013, Recombination-mediated genetic engineering of Plasmodium berghei DNA, Malaria, 923, 127

Howard et al., 2013, Synthesis of customized petroleum-replica fuel molecules by targeted modification of free fatty acid pools in Escherichia coli, PNAS, 110, 7636

Kim et al., 2013, Cloning and characterization of a putative UDP-rhamnose synthase 1 from Populus euramericana Guinier, J Plant Biol, 56, 7

Heinrich et al., 2013, From waste to plastic: Synthesis of pPpoly(3-Hydroxypropionate) in Shimwellia blattae, AEM, 79, 3582

Tan et al., 2013, Activating phosphoenolpyruvate carboxylase and phosphoenolpyruvate carboxykinase in combination for improving succinate production, AEM, doi: 10.1128/AEM.00826-13

Kim et al., 2013, Biosynthesis of bioactive O-methylated flavonoids in Escherichia coli, AMB, 97, 7195

Arbel-Goren et al., 2013, Effects of post-transcriptional regulation on phenotypic noise in Escherichia coli, Nucleic Acid Res, 41, 4825

Wang et al., 2013, Engineered heterologous FPP synthases-mediated Z,E-FPP synthesis in E. coli, Metabolic Eng, 18, 53

Kim et al., 2013, Production of hydroxycinnamoyl-shikimates and chlorogenic acid in Escherichia coli: production of hydroxycinnamic acid conjugates, Microbiol Cell Factories, 12, 15

Xiao et al., 2013, Engineering Escherichia coli to convert acetic acid to free fatty acids, Biochem Engineering J, 76, 60

Cimini et al., 2013, Homologous overexpression of rfaH in E. coli K4 improves the production of chondroitin-like capsular polysaccharide, Microbial Cell Factories, 12, 46

2013/2014: Other approaches

Fried et al., 2013, Identification of a target gene and activating Stimulus for the YpdA/YpdB histidine kinase/response regulator system in Escherichia coli, J Bacteriol, 195, 807

Sivakamasundari et al., 2013, Pax1EGFP: New wildtype and mutant EGFP mouse lines for molecular and fate mapping studies, Genesis, 51, 420

Huong et al., 2013, In vivo analysis of aicda gene regulation: A critical balance between upstream enhancers and intronic silencers governs appropriate expression, PlosOne 8, e61433

Katsumata et al., 2013, Bioluminescence imaging of β cells and intrahepatic insulin gene activity under normal and pathological conditions, PlosOne, 8, e60411

Miyazaki et al., 2013, Galnt3 deficiency disrupts acrosome formation and leads to oligoasthenoteratozoospermia, Histochem & Cell Biol, 139, 339

Zhou et al., 2013, An essential function for the ATR-Activation-Domain (AAD) of TopBP1 in mouse development and cellular senescence, PlosGeneteics, 9, e1003702

Spatz et al., 2013, Expression of chicken parvovirus VP2 in chicken embryo fibroblasts requires codon optimization for production of naked DNA and vectored meleagrid herpesvirus type 1 vaccines, Virus Genes, 47, 259

Webber et al., 2013, The relationship between long-range chromatin occupancy and polymerization of the Drosophila ETS family transcriptional repressor Yan, Genetics, 193, 633

Sugimoto et al., 2013, Generation and characterization of ScxCre transgenic mice, Genesis, 51, 275

Nagase et al., 2013, Deiodinase 2 upregulation demonstrated in osteoarthritis patients cartilage causes cartilage destruction in tissue-specific transgenic rats, Osteoarthritis and Cartilage, 21, 514

Thomas et al., 2013, Piwi induces piRNA-guided transcriptional silencing and establishment of a repressive chromatin state, Genes&Dev, 27, 390

Schraml et al., 2013, Genetic tracing via DNGR-1 expression history defines dendritic cells as a hematopoietic lineage, Cell, 154, 843

Tanaka et al., 2013, Astrocytic Ca2 signals are required for the functional integrity of tripartite synapses, Molecular Brain, 6, 6

Akieda-Asai et al., 2013, Involvement of guanylin and GC-C in rat mesenteric macrophages in resistance to a high-fat diet, J Lip Res, 54, 85

Charleswort, 2013, Plant sex chromosome evolution, J Experi Botany, 64, 405

Chien et al., 2013, Misregulated progesterone secretion and impaired pregnancy in Cyp11a1 transgenic mice, Biol of Reproduction, doi:10.1095/biolreprod.113.110833

Manu et al., 2013, Sex-specific pattern formation during early Drosophila development, Genetics, 194, 163

Ogiwara et al., 2013, Nav1.1 haploinsufficiency in excitatory neurons ameliorates seizure-associated sudden death in a mouse model of Dravet syndrome, Hum Mol Genetics, 22, 4784

Sato et al., 2013, Fluoroquinolone resistance mechanisms in an Escherichia coli isolate, HUE1, without quinolone resistance-determining region mutations, Front Microbiol, 4, 125

Tanaka et al., 2013, Epigenetic Thpok silencing limits the time window to choose CD4 helper-lineage fate in the thymus, EMBO J, 32, 1183

Wang et al., 2013, A molecular genetic approach to uncovering the differential functions of dopamine D2 receptor isoforms, Dopamine: Methods in Mol Biol, 964

Fukuzawa et al., 2013, SGLT5 reabsorbs fructose in the kidney but its deficiency paradoxically exacerbates hepatic steatosis induced by fructose, PlosOne, 8, e56681

Morales et al., 2013, Utx is required for proper induction of ectoderm and mesoderm during differentiation of embryonic stem cells, PlosOne, 8, e60020

Murata et al., 2013, Contribution of myocyte enhancer factor 2 family transcription factors to BZLF1 expression in Epstein-Barr virus reactivation from latency, J Virology, 87, 10148

Liu et al., 2013, Direct and indirect roles of Fgf3 and Fgf10 in innervation and vascularisation of the vertebrate hypothalamic neurohypophysis, Development, 140, 1111

Klose et al., 2013, Fosmid-based structure-function analysis reveals functionally distinct domains in the cytoplasmic domain of Drosophila crumbs, G3, 3, 153

Delenda et al., 2013, Bio-applications derived from site-directed genome modification technologies, site-directed insertion of transgenes, Topics in Current Genetics, 23

Koayashi et al., 2013, Genetic dissection of medial habenula–interpeduncular nucleus pathway function in mice, Frot Behav Neurosci, 7, 17

Matkovic et al., 2013, The Bruchpilot cytomatrix determines the size of the readily releasable pool of synaptic vesicles, JCB, 202, 667

Hu et al., 2013, VirD: A virion display array for profiling functional membrane proteins, Anal Chem, 85, 8046


Wang et al., 2006, An improved recombineering approach by adding RecA to lambda Red recombination, Mol Biotechnol, 32, 43

Wenzel et al., 2005, Heterologous expression of a myxobacterial natural products assembly line in pseudomonads via red/ET recombineering, Chem Biol, 12, 349

Glaser et al., 2005, Current issues in mouse genome engineering, Nat Genet, 37, 1187

Vetcher et al., 2005 Rapid engineering of the geldanamycin biosynthesis pathway by Red/ET recombination and gene complementation, Appl Environ Microbiol, 71, 1829

Ikeya et al., 2005, Gene disruption/knock-in analysis of mONT3: Vector construction by employing both in vivo and in vitro recombinations, Int J Dev Biol, 49, 807

Testa et al., 2004, A reliable lacZ expression reporter cassette for multipurpose, knockout-first alleles. Genesis, 38, 151

Testa et al., 2003, Engineering the mouse genome with bacterial artificial chromosomes to create multipurpose alleles. Nat Biotechnol, 21, 443

Zhang et al., 2003, Phage annealing proteins promote oligonucleotide-directed mutagenesis in Escherichia coli and mouse ES cells, BMC Mol Biol, 4, 1

Muyrers et al., 2002, Introducing Red/ET recombination: DNA engineering for the 21st Century, Gene Cloning & Expression Technologies, Biotechniques

Shimshek et al., 2002, Codon-Improved Cre recombinase (iCre) expression in the mouse, Genesis 32, 19

Voziyanov et al., 2002, A dual reporter screening system identifies the amino acid at position 82 in Flp site-specific recombinase as a determinant for target specificity, Nucleic Acids Res, 30, 1656

Muyrers et al., 2001, Recombinogenic engineering: New options for manipulating DNA Trends in Biochemical Sciences, 26, 325

Schaft et al., 2001, Efficient FLP recombination in mouse ES cells and oocytes, Genesis, 31, 6

Buchholz et al., 2001, LoxP-directed cloning: Use of Cre recombinase as a universal restriction enzyme, Biotechniques, 31, 906

Muyrers et al., 2000, ET cloning: Think recombination first, Genetic Engineering, Principles and Methods, 22, 77

Muyrers et al., 2000, Point mutation of bacterial artificial chromosomes by ET recombination, EMBO Reports, 1, 239

Hill et al., 2000, BAC trimming minimizing clone overlaps, Genomics, 64, 111

Muyrers et al., 2000, RecE/RecT and Reda/Redb initiate double stranded break repair by specifically interacting with their respective partners, Genes and Development, 14, 1971

Zhang, et al., 2000, DNA cloning by homologous recombination in Escherichia coli, Nature Biotechnology 18, 1314

Rodriguez et al., 2000, High-efficiency deleter mice show that FLPe is an alternative to Cre-loxP, Nat Genet. 25, 139

Testa et al., 2000, Creating a transloxation: Interchromosomal translocations in mice, EMBO Reports, 1, 120

Muyrers et al., 1999, Rapid modification of Bacterial Artificial Chromosomes by ET-recombination, Nucleic Acids Res, 27, 1555

Angrand et al., 1999, Simplified generation of targeting constructs using ET recombination, Nucleic Acids Res, 27, e16

Narayanan et al., 1999, Efficient and precise engineering of a 200 kb beta-globin human/bacterial artificial chromosome in E. coli DH10B using an inducible homologous recombination system, Gene Ther, 6, 442

Ringrose et al., 1999, Quantitative comparison of DNA looping in vitro and in vivo: Chromatin increases effective DNA flexibility at short distances, EMBO J, 23, 6630

Zhang et al., 1998, A new logic for DNA engineering using recombination in E. coli, Nature Genetics, 20, 123

Schwenk et al., 1998, Temporally and spatially regulated somatic mutagenesis in mice, Nucleic Acids Res, 26, 1427

Angrand et al., 1998, Inducible transgene expression based on regulated recombination, Nucleic Acids Res, 26, 3263

Ringrose et al., 1998, Comparative kinetic analysis of Flp and Cre recombinases: Mathematical models for DNA binding and recombination, J Mol Biol, 284, 363

Ringrose et al., 1997, The Kw recombinase, an integrase from Kluyveromyces waltii, Eur J Biochem, 248, 903

Buchholz et al., 1996, A simple assay to verify the functionality of Cre and FLP recombination targets in genomic manipulation constructs, Nucleic Acids Res, 24, 3118

Buchholz et al., 1996, Different thermostabilities of FLP and Cre recombinases: Implications for applied site specific recombination, Nucleic Acids Res, 24, 4256

Logie et al., 1995, Ligand-regulated site-specific recombination, PNAS, 92, 5940

If you cannot find the answer to your problem below then please contact us or telephone 01954 210 200

Quick and Easy BAC Modification

The Quick and Easy BAC Modification kit is designed to modify any type of bacterial artificial chromosomes (BACs) within 1-2 weeks by using a kanamycin/neomycin cassette

Gene Bridges

Kits including Red/ET products will require a signed and accepted sub-license


Please contact us for details or download the acadmic version here.

If you cannot find the answer to your problem below then please contact us or telephone 01954 210 200

Quick and Easy BAC Modification

The Quick and Easy BAC Modification kit is designed to modify any type of bacterial artificial chromosomes (BACs) within 1-2 weeks by using a kanamycin/neomycin cassette

Gene Bridges


  • Simple BAC modification like insertion of a selection marker, deletion of a fragment with a selection marker gene, insertion of a function fragment together with selection marker gene
  • This kit can also be applied on bacterial chromosome modification


  • Fast and reliable cloning of large DNA molecules
  • Precise DNA modification at position of choice
  • Independent of restriction sites
  • No size limitation

If you cannot find the answer to your problem below then please contact us or telephone 01954 210 200