Cell Signaling Kinases T4 Polynucleotide Kinase Product T4 Polynucleotide Kinase Available Separately T4 PNK Buffer Pack For Laboratory Use. Description: T4 Polynucleotide Kinase catalyzes the transfer of the γ-phosphate from ATP to the 5´-terminus of polynucleotides or to mononucleotides bearing a 5´-hydroxyl group. The enzyme, purified from recombinant E. coli, may be used to phosphorylate RNA, DNA and synthetic oligonucleotides prior to subsequent manipulations such as ligation. Features: • May Be Heat-Inactivated: T4 Polynucleotide Kinase may be inactivated by heating at 68°C for 10 minutes. • Provided with 10X Reaction Buffer: 700mM Tris-HCl (pH 7.6 at 25°C), 100mM MgCl2, 50mM DTT. • Blue/White Cloning Qualified: Promega’s blue/white cloning assay provides a higher level of quality control for enzymes used in cloning applications. • Choose Your Configuration: Learn more about our custom options for this product at: www.promega.com/myway/ Storage Conditions: Store at –20°C. Protocol T4 Polynucleotide Kinase Technical Bulletin Nucleases Exonuclease III Product Exonuclease III For Laboratory Use. Description: Exonuclease III is a 3´→5´ exonuclease specific for doublestranded DNA. The enzyme catalyzes the stepwise removal of mononucleotides starting from a 3´-OH at nicks, blunt ends, recessed ends and 3´-overhangs of less than 4 bases, yielding nucleoside 5´-phosphates. Exonuclease III will degrade DNA from 3´-phosphate ends due to its intrinsic 3´-phosphatase activity. In addition, the enzyme has apurinic endonuclease activity and ribonuclease H activity. Exonuclease III is used in conjunction with S1 nuclease for unidirectional deletion of sequences from the termini of DNA fragments. Features: • Flexible: Control deletion rate by varying incubation temperature. • May Be Heat-Inactivated: Exonuclease III may be inactivated by heating to 75°C for 10 minutes. • Provided with 10X Reaction Buffer: 660mM Tris-HCl (pH 8.0 at 25°C), 6.6mM MgCl2. Storage Conditions: Store at –20°C. Protocol Exonuclease III Protocol Part# 9PIM181 Size Conc. Cat.# Price (Fr) 5,000 u 150–200 u/µl M1811 119.00 25,000 u 150–200 u/µl M1815 417.00 Part# TB519 Size Conc. Cat.# Price (Fr) 100 u 5–10 u/µl M4101 89.00 1,000 u 5–10 u/µl M4103 623.00 Size Cat.# Price (Fr) 1.5 ml C1313 57.00 Erase-a-Base™ System Product Erase-a-Base™ System (minus vectors & bacterial strain) For Research Use Only. Not for Use in Diagnostic Procedures. Description: The Erase-a-Base™ System is designed for the rapid construction of plasmid or M13 subclones containing progressive unidirectional deletions of any inserted DNA. The system is based on the procedure developed by Henikoff, in which exonuclease III (Exo III) is used to specifically digest insert DNA from a 5´ protruding or blunt end restriction site. The adjacent sequencing primer binding site is protected from digestion by a 4-base 3´ overhang restriction site or by an α-phosphorothioate-filled end. Features: • Rapid: Construction of nested deletions from plasmid or M13 clones are rapid. Construction is complete in a few hours. • Efficient: Produce deletion sets spanning several kilobases. Storage Conditions: Store at –20°C. Protocol Erase-a-Base® System Technical Manual Mung Bean Nuclease Product Mung Bean Nuclease Size For Research Use Only. Not for Use in Diagnostic Procedures. Description: Mung Bean Nuclease catalyzes the degradation of singlestranded DNA and RNA endonucleolytically to yield 5´-phosphoryl-terminated products. While the nuclease prefers ssDNA over dsDNA by 30,000-fold, at very high concentrations the enzyme degrades double-stranded DNA from both ends. Mung Bean Nuclease has been used for transcript mapping studies, for flushing staggered ends and for the separation of cDNA strands after synthesis with reverse transcriptase and DNA Polymerase I. Features: • Provided with 10X Reaction Buffer: 300mM sodium acetate (pH 5.0 at 15°C), 500mM NaCl, 10mM ZnCl2. Storage Conditions: Store at –20°C. Protocol Mung Bean Nuclease Protocol Part# 9PIM431 Conc. Cat.# Price (Fr) 2,000 u 50–100 u/µl M4311 143.00 Part# TM006 Size Cat.# Price (Fr) 1 system E5750 830.00 4 For complete and up-to-date product information visit: www.promega.com/catalog 93 Cloning and DNA Markers Pagina 96
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