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.

Alkaline Phosphatase, shrimp
Shrimp Alkaline Phosphatase (SAP) catalyzes the dephosphorylation of 5’ phosphates from DNA and RNA. SAP is equally efficient on either 5’-protruding, 5’-recessive and blunt ends. Unlike calf intestinal phosphatase, SAP is completely and irreversibly inactivated by heat treatment for 15 min at 65°C. Thus the whole procedure including restriction enzyme digestion, dephosphorylation, enzyme inactivation, and ligation with the Rapid DNA Ligation Kit is performed in one single tube.

Expand Cloning Kit
The Expand Cloning Kit provides all reagents for a fast and efficient method for cloning of large PCR fragments in a size range of 7-36 kb. The kit also provides reagents to perform a onestep reaction (when necessary), where the PCR fragments are polished and subsequently phosphorylated. The Expand vectors are supplied linearized and dephosphorylated. The Expand Cloning Kit is optimized for cloning of long PCR fragments, e.g., generated with the Expand Long Template PCR System.

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 Ligation Kit
The Rapid DNA Ligation Kit enables ligation of sticky end or blunt-end DNA fragments in just 5 min at 15°C - 25°C. Depending on DNA concentration, either circular (low DNA concentration) or concatemeric (high DNA concentration) ligation products are formed. All necessary reagents required for ligation are provided. There is no need to prepare buffers, or to add ATP or Mg²⁺.

Shrimp Alkaline Phosphatase
Shrimp Alkaline Phosphatase (SAP) catalyzes the dephosphorylation of 5’ phosphates from DNA and RNA. SAP is equally efficient on either 5’-protruding, 5’-recessive and blunt ends. Unlike calf intestinal phosphatase, SAP is completely and irreversibly inactivated by heat treatment for 15 min at 65°C. Thus the whole procedure including restriction enzyme digestion, dephosphorylation, enzyme inactivation, and ligation with the Rapid DNA Ligation Kit is performed in one single tube.

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.