• Synthesis of dsRNA from the 3' end of a ssRNA template
• Highly processive primer independent RNA synthesis
• Uses a short 9 nt initiator sequence
Description:
The phi6 RNA polymerase is a modified version of the P2 protein of bacteriophage phi6. It is an RNA-dependent RNA polymerase (RdRp) which synthesizes a full length complementary strand of an RNA initiating from the 3´ end resulting in dsRNA. The polymerase does not require any oligonucleotide primer for initiation. phi6 RdRp is a highly processive enzyme that can use either single or double stranded RNA template. The template strand of RNA requires the sequence 5´ NNUUUUUUUUCC 3´ for optimal initiation (1-3).
Source:
An E. coli strain that carries a modified P2 gene from bacteriophage phi6.
Reaction Conditions:
1X phi6 RdRp Reaction Buffer
Supplemented with 1.5 mM MnCl2
Incubate at 32°C.
1X phi6 RdRp Reaction Buffer:
50 mM Tris-HCl
50 mM Ammonium Acetate
pH 8.9 @ 25°C
Unit Definition:
One unit is defined as the amount of enzyme that will incorporate 1 nmol of UTP into an acid-insoluble material in 20 minutes at 32°C.
Unit Assay Conditions:
1 X phi6 RdRp Reaction Buffer, 1.5 mM MnCl2, 10% DMSO, 1 mM UTP, 1 µg poly (rA) and 1 µCi 3H-UTP in 30 µl.
Concentration:
1,000 units/ml
Storage Conditions:
50 mM Tris-HCl
100 mM NaCl
0.1 mM EDTA
50% Glycerol
0.1% Triton X-100
pH 8.0 @ 25°C
原厂资料：
• Synthesis of dsRNA for RNAi experiments
• Synthesis of dsRNA from the 3' end of a ssRNA template
• Highly processive primer independent RNA synthesis
• Uses a short 9 nt initiator sequence
Description:
The phi6 RNA polymerase is a modified version of the P2 protein of bacteriophage phi6. It is an RNA-dependent RNA polymerase (RdRp) which synthesizes a full length complementary strand of an RNA initiating from the 3´ end resulting in dsRNA. The polymerase does not require any oligonucleotide primer for initiation. phi6 RdRp is a highly processive enzyme that can use either single or double stranded RNA template. The template strand of RNA requires the sequence 5´ NNUUUUUUUUCC 3´ for optimal initiation (1-3).
Source:
An E. coli strain that carries a modified P2 gene from bacteriophage phi6.
Reaction Conditions:
1X phi6 RdRp Reaction Buffer
Supplemented with 1.5 mM MnCl2
Incubate at 32°C.
1X phi6 RdRp Reaction Buffer:
50 mM Tris-HCl
50 mM Ammonium Acetate
pH 8.9 @ 25°C
Unit Definition:
One unit is defined as the amount of enzyme that will incorporate 1 nmol of UTP into an acid-insoluble material in 20 minutes at 32°C.
Unit Assay Conditions:
1 X phi6 RdRp Reaction Buffer, 1.5 mM MnCl2, 10% DMSO, 1 mM UTP, 1 µg poly (rA) and 1 µCi 3H-UTP in 30 µl.
Concentration:
1,000 units/ml
Storage Conditions:
50 mM Tris-HCl
100 mM NaCl
0.1 mM EDTA
50% Glycerol
0.1% Triton X-100
pH 8.0 @ 25°C
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