PCR cloning kits
Alkaline Phosphatase,
From Calf Intestine
Alkaline Phosphatase hydrolyzes 5'-terminal monophosphate
groups of DNA and RNA and, 5'-di- and triphosphate groups of RNA.
Polynucleotide
Kinase
Polynucleotide Kinase catalyzes the transfer of the terminal phosphate group
of ATP to the 5’-hydroxylated terminus of DNA or RNA. It exhibits 3'-phosphatase
activity, which is exhibited at low pH. The enzyme also catalyzes the exchange
of 5’-terminal phosphate groups.
PCR Cloning Kit
(blunt-end)
The PCR Cloning Kit (blunt-end) offers a fast, efficient, and convenient
method to clone small DNA fragments (<1.5 kb) as well as larger DNA fragments
up to 10 kb. Blunt-ended DNA fragments (e.g., PCR fragments) can be
used directly for rapid ligation (5 min) to achieve high cloning efficiencies.
Any blunt-ended DNA fragment may be used directly for cloning purposes, without
pretreating the ends (i.e., purification or polishing).
Rapid DNA Dephos and Ligation
Kit
Upgrade from the Rapid DNA Ligation Kit and choose the new Rapid DNA Dephos & Ligation Kit for fast and efficient dephosphorylation and ligation of sticky- or blunt-end DNA fragments. This kit contains the new recombinant rAPid Alkaline Phosphatase, the tool of choice for dephosphorylation of 5' phospahates from nucleic acids.
T4 DNA Ligase
T4 DNA Ligase catalyzes the formation of phosphodiester bonds between neighbouring
3'-hydroxyl and 5'-phosphate ends in double-stranded DNA. Single-stranded nicks
in double-stranded DNA are also closed.
T4 DNA Polymerase
T4 DNA Polymerase is a DNA dependent DNA polymerase that catalyzes the polymerization
of deoxynucleoside-5´-triphosphates to the hydroxyl termini of recessive
ends. Blunt-ended DNA cannot serve as a template for the polymerization reaction.
T4 DNA Polymerase requires DNA with 5´- protruding ends and a high concentration
of dNTP’s for polymerization. The enzyme carries an extremely active 3´
- 5´-exonuclease that shows a strong specificity for ssDNA and lacks a
5´ - 3´-exonuclease activity. Therefore nicked duplex DNA cannot
serve simultaneously as a template and primer for polymerization. The addition
of T4 Gene 32 Protein facilitates strand displacement, and therefore allows
T4 DNA polymerase to replicate the nicked duplex.
T4 Polynucleotide
Kinase
T4 Polynucleotide Kinase, 3’-phosphatase free, catalyzes the transfer
of the terminal phosphatase group of ATP to the 5’-hydroxylated terminus
of DNA or RNA. It also exchanges 5’-terminal phosphate groups. This phage
derivative lacks 3’-phosphatase activity.
T4 RNA Ligase
Bacteriophage T4 RNA Ligase catalyzes the ATP-dependent covalent joining of
a 5´-phosphoryl-terminated nucleic acid donor to a 3´-hydroxyl-terminated
nucleic acid acceptor. Substrates are single stranded DNA and RNA as well as
mononucleosides and 3´, 5´-biphosphates (pNp). An oligonucleotide
that has both a 5´-phosphate and a 3´-hydroxyl can also serve as
either an acceptor or a donor, yielding either circular or multimeric products.
Terminal Transferase
Terminal Transferase catalyzes the template independent addition of deoxy- and
dideoxynucleoside triphosphates to the 3´OH ends of double- and single-stranded
DNA fragments and oligonucleotides. Terminal Transferase incorporates digoxigenin-,
biotin-, and fluorochrome-labeled deoxy- and dideoxynucleoside triphosphates
as well as radioactive labeled deoxy- and dideoxynucleoside triphosphates.