Invitrogen topo ta cloning kit for sequencing manual

TOPO TA Cloning® Kit for Sequencing supplied with the PureLink™ Quick Plasmid Miniprep (cat. no.K) is shipped with an additional box containing reagents for plasmid purification (Box 3). Store Box 1 at °C, Box 2 at °C, and Box 3 at room temperature. Types of Kits TOPO TA Cloning® Kits for Sequencing are available with either Mach1™-T1R. The TOPO ® TA Cloning ® Kits for Sequencing provide a highly efficient, 5 minute, one-step cloning strategy ("TOPO ® Cloning") for the direct insertion of Taq polymerase-amplified PCR products into a plasmid vector for sequencing. No ligase, post-PCR procedures, or PCR primers containing specific sequences are required. Use the Invitrogen TOPO TA Cloning kit. Please read the instruction manual carefully before starting. Invitrogen has changed the kit several times in the last year. The latest manuals are versions J and K as of 9/ Please note that the kits no longer contain b-mercaptoethanol or stop solution but now contain a salt solution which is added to the cloning reaction. The TOPO vector has an overhanging 3' T.

Kit includes everything necessary to clone and select recombinant vectors containing your PCR fragment of choice. pCR4-TOPO TA Vector: Optimized for Sequencing We have removed much of the multiple cloning site from the pCR4-TOPO TA vector to shorten the distance between sequencing primer sites and the insert site to as little as 33 bp. This. Upon arrival, store the competent cells (in the large exterior box) at °C, the components in the small interior box at °C and the NEB™ beta/Stable Outgrowth Medium at room temperature or 4°C. Product Notes. The NEB PCR Cloning Kit contains a sufficient supply of materials to perform 20 x 10 μl cloning reactions. After the 20 minute incubation, place the ligation reaction on ice. Add 2 μl cloning reaction into vial of chemically competent E coli and mix gently (don't mix by pipetting up and down). Incubate on ice 20 minutes. Heat-shock cells by putting the microfuge tube from Step 3 in the 42 °C waterbath or heat block for exactly 2 minutes.

Introduction pBAD TOPO® TA Expression Kit provides a highly efficient, 5-minute, one-step cloning strategy ("TOPO® Cloning") for the direct insertion of Taq polymerase-amplified PCR products into a plasmid vector for regulated expression in E. coli. No ligase, post-PCR procedures, or PCR primers containing specific sequences are required. The TOPO® TA Cloning® Kits for Sequencing provide a highly efficient, 5-minute, one-step cloning strategy ('TOPO® Cloning') for the direct insertion of Taq polymerase–amplified PCR products into a plasmid vector for sequencing. Each kit uses the pCR™4-TOPO® TA vector with specially designed sequencing primer sites that return more insert sequence and less vector sequence from each reaction, and are available with a variety of competent cells, or no competent cells, depending on your. Introduction The pENTR™ Directional TOPO® Cloning Kits utilize a highly efficient, 5-minute cloning strategy ("TOPO® Cloning") to directionally clone a blunt-end PCR product into a vector for entry into the Gateway ® System or the MultiSite Gateway System available from Invitrogen. Blunt-end PCR products clone directionally at greater.