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商品編號


M0310S-5,000 units



品牌


紐英倫(NEB)



公司


New England Biolabs



公司分類(lèi)


Nucleic Acid Purification



Concentration

10,000 units/ml


Size

5,000 units

商品信息

Description:
The p19 siRNA Binding Protein (19 kDa) from the Carnation Italian Ringspot Virus (CIRV) plant binds siRNAs with nanomolar affinity(1). The dimeric protein binds 21 base siRNAs with a 2 base 3? extension and a 5?phosphate. The protein binds RNA in a size dependent and sequence independent manner. If the siRNAs are 4 bases longer, the affinity for the protein is reduced about 100-fold (2). When p19 siRNA Binding Protein is expressed in plants it suppresses RNA interference (3).
Figure 1:
Size specific binding of siRNA by p19 siRNA Binding Protein. Three phosphorylated dsRNAs of 17,21 and 25 bases (30 ng each band) were mixed with increasing amount of p19 siRNA Binding Protein (0-3 ?g) in a 20 ?l reaction, and incubated at room temperature for 1 hour. The reaction was analyzed on a 20% polyacrylamide gel stained with ethidium bromide.
Marker
M is the siRNA
Marker
(
NEB
#N2101 ).
Highlights
For the isolation of siRNA
Isolated from a recombinant source
Product Source
p19 siRNA Binding Protein is cloned and expressed in?
E. coli
?as a fusion protein with an amino terminal MBP (maltose binding protein) and a carboxy terminal?C
BD
(chitin binding domain).
Reagents Supplied
The following reagents are supplied with this product:
Store at (°C)
Concentration
p19 siRNA Binding Buffer?(200?μl)
2X
Notes:
p19 siRNA Binding Protein can selectively bind siRNAs in the presence of a 2,000 fold excess of other RNAs.1X p19 siRNA Binding Buffer:20 mM Tris-HCl, 100 mM NaCl, 1 mM EDTA, 1 mM TCEP, 0.02% Tween-20 (pH 7.0 @ 25°C)p19 siRNA Binding Protein does not bind ssRNA.Double-stranded 21 bp RNA with a 2-base overhang and a 5? phosphate binds the most efficiently. Double-stranded miRNAs with mismatched base pairs will bind with lower affinity.Due to the difference in molecular weight between p19 siRNA Binding Protein and siRNA, a 10 to 20-fold excess of p19 siRNA Binding Protein is needed.TCEP (Tris 2-carboxyethyl Phosphine) can be replaced with DTT (dithiotreitol) in the required solutions. However, if DTT is used, it needs to be added separately into the reaction.