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Invitrogen™ Bac-to-Bac™ N-His TOPO™ Cloning Kit Non-distribution product as customer accommodation.

Bac-to-Bac™ N-His TOPO™ Cloning Kit

Manufacturer:  Invitrogen™ A11099

Catalog No. A11099


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Description

Description

Bac-to-Bac™ Baculovirus Expression is an efficient method for producing baculovirus for high-level protein expression in insect cells. It relies on generation of recombinant virus by site-specific transposition in E. coli rather than homologous recombination in insect cells. New: pFastBacTOPO™ vectors!

Reliable and fast protein expression
The Bac-to-Bac™ Baculovirus Expression System is faster and easier than traditional baculovirus expression, and it maintains high levels of protein expression. It is based on site-specific transposition in E. coli rather than homologous recombination in insect cells. The Bac-to-Bac™ Baculvirus Expression System is highly regarded in academic Literature. At least 80 citations every year since 2000!

Traditional baculovirus systems require purification and amplification of an initial low-titer viral supernatant. This requires a time-consuming plaque assay. Bac-to-Bac™'s pFastBac™ vector, however, recombines with the parent bacmid in DH10Bac™ E. coli competent cells to form an expression bacmid. Transfect the bacmid into insect cells for fast production of a high titer of pure recombinant baculovirus particles in the very first transfection. You'll save weeks of precious time. Collect a pure P2 baculovirus stock on Day 10 without the necessity of tedious plaques assay (See Figure 1)

High expression, easy screening
pFastBac™ uses the strong polyhedrin promoter to generate high levels of expression in a variety of insect cell lines such as Sf9, Sf21, and High Five™ cells.

New additions:
Bac-to-Bac™'s pFastBac vectors are now available as pFastBacTOPO™ vectors too. pFastBacTOPO™ vectors are Blunt™ TOPO™ vectors and are supplied with Mach1™-T1R E. coli for easy cloning and pFastBac⁄GOI propagation. The new vectors enable you to:

  • Amplify your gene of interest (GOI) with a PCR enzyme of highest fidelity such as Accuprime™ Pfx SuperMix.

  • TOPO™ clone the blunt-end PCR product into the new vector in only 5 minutes!

  • Visualize colonies 8 hours after plating on ampicillin selective plates because Mach1™-T1R E. coli cells have a faster doubling time compared to other standard cloning strains.

  • With the Bac-to-Bac™ N-His TOPO™ cloning or expression kit, you can produce a N-terminal His-fusion protein with a TEV cleavage site to purify with nickel-chelating resin (Invitrogen’s ProBond™ Purification System) and generate a native protein with the aid of Invitrogen’s popular AcTEV™ Protease.

    The new vectors combine the TOPO™ cloning technology with the highly regarded Bac-to-Bac™ baculovirus expression features to enable easy cloning and high-level protein expression.
    The pFastBacTOPO™ vectors are offered as cloning and expression kits.
  • Specifications

    Specifications

    Ampicillin (AmpR), gentamicin (GmR)
    Protein expression, viral vaccine production, protein biology, protein expression, insect expression, high-level protein production in insect cells, baculovirus production and titering, bioproduction, viral vaccine production
    Vector kit for 20 topo cloning reactions: pFastBac/NT-TOPO™ Vector containing the N-terminal His-Tag and TEV cleavage site, pFastBac/NT-Gus control plasmid (Control expression vector), 10x PCR, dNTP Mix, salt solution, sterile water, control PCR template, polyhedrin forward primer, SV40 pA reverse primer. Kit containing competent cells (20 reactions): One Shot™ Mach1-T1R chemically competent E. coli vector kit: store at –20°C
    Polyhedrin forward, SV40 pA reverse
    20 rxn(s)
    –30 to –10°C
    Bac-to-Bac™ N-His TOPO™ Cloning Kit, 1 Kit, 20 rxns, High Five™, Sf21, Sf9 Cell Line, Baculovirus Cell-Based Expression, Polyhedrin Promoter, His Tag Protein, Blunt TOPO™ Cloning Method, pFastBac/NT-TOPO Vector, Six Months Shelf Life, –30 to –10°C Storage Temperature, Used for Protein Expression and Viral Vaccine Production
    Liquid
    20 rxn(s)
    Bac-to-Bac™ N-His TOPO™ Cloning Kit
    5'-AAATGATAACCATCTCGC-3', 5'-GGTATGGCTGATTATGATC-3'
    pFastBac/NT-TOPO
    Safety and Handling

    Safety and Handling

    ShelfLife : 6 Month(s)

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