This package includes the CloneJET PCR cloning kit (20 or 40 reactions) and DH10B competent cells, which are optimized for Used in conjunction with the kit, the ligation product can be used directly for transformation.

CloneJET PCR Cloning Kit
The Thermo Scientific CloneJET PCR Cloning Kit is an advanced positive selection system for PCR generated with any thermostable DNA polymerase Efficient cloning of products. Any blunt-ended or sticky-ended DNA fragment can also be cloned. This kit can be used with both phosphorylated and non-phosphorylated DNA fragments. Ligate to the included positive selection vector in just five minutes and obtain over 99% recombinant clones. The blunt-ended PCR product generated by the proofreading enzyme is directly inserted into the cloning vector.

PCR products generated by non-proofing DNA polymerase or DNA polymerase mixtures need to be treated into blunt ends using the thermostable DNA blunt-endase provided in the kit within a five-minute reaction time before ligation. DH10B competent cells are optimized for use with the CloneJET PCR cloning kit and can be directly transformed with ligation products.

CloneJET PCR cloning kit contains the new ready-to-use positive selection cloning vector pJET1.2/blunt. The vector contains a lethal restriction enzyme gene that is destroyed after the DNA insert is inserted into the cloning site. Therefore, only bacterial cells containing the recombinant plasmid can form colonies. The recirculated pJET1.2/blunt vector molecule without the insert expresses a lethal restriction enzyme that kills the host E. coli cells upon transformation. This positive screening greatly speeds up the colony screening process and saves the additional cost required for blue and white spot screening.

•Fast — PCR cloning in just five minutes
• Extremely efficient — >99% positive clones
• No cloning background — Positive selection vector
• Universal — Suitable for blunt-end or sticky-end cloning
• Economical — No need Perform expensive blue/white screening

Applications include:
• Cloning of blunt-ended or 3' dA tailed PCR products up to 10 kb
• Cloning of DNA generated by restriction enzymes Fragments
• Sequencing of cloned DNA
• In vitro and in vivo transcription of cloned inserts using the T7 promoter

Learn more about CloneJET PCR Cloning Kit›

DH10B Competent Cells
Thermo Scientific DH10B Competent Cells are highly efficient chemically competent E. coli cells. This DH10B strain is suitable for cloning DNA containing methylcytosine and methyladenine, and can efficiently clone prokaryotic and eukaryotic genomic DNA. These cells are suitable for cDNA library construction using plasmid-derived vectors. Genotype: F– mcrA Δ(mrr-hsdRMS-mcrBC) φ80lacZΔM15 ΔlacX74 recA1 endA1 araD139 Δ (ara-leu)7697 galU galK λ– rpsL(StrR) nupG.

•High transformation efficiency: >1 x 109 cfu/µg pUC19 DNA
• Suitable for routine and high-throughput cloning applications
• Convenient, two reactions per vial
>• SOC containing external growth medium

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