服務(wù)熱線(xiàn)：
Biohelix/ET SSB/H0221S/50?g
克隆和表達
商品編號
H0221S
品牌
Biohelix
公司
BIO-HELIX Co., LTD.
公司分類(lèi)
Primer Navigator? series
Size
50?g
Concentration
0.5 mg/ml
商品信息
Description:
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Single-Strand Binding Protein (SSB) binds to single-stranded DNA
with high affinity and also binds to RNA and double-stranded DNA
with lower affinity (1).
In vivo
, it st
ABI
lizes transiently
formed ssDNA and plays an important role in DNA replication, recombination
and repair (2).
In vitro
, SSB proteins have been used to
dest
ABI
lize secondary structures in DNA and to increase the processivity
of DNA polymerases in several molecular
BIOLOG
y applications: SSBs
improve the yield and efficiency of reverse transcription reactions
during RT-PCR as well as increase the yield of PCR products (3-9).
ET SSB (Extreme
Thermo
stable SSB)
is a single-stranded DNA binding protein isolated from a hyper
Thermo
philic
microorganism, and it is a flagship enhancer of the Primer Navigator
product series. It remains fully active after incubation at 95?C
for 60 min. Due to the extreme
Thermo
st
ABI
lity,
ET
SSB
can be used in applications that require extremely
high temperature conditions, such as nucleic acid amplification
and sequencing.
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Source:
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Purified from an
E. coli
strain that overexpresses the ssb gene isolated from a hyper
Thermo
philic
microorganism.
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Applications:
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Improve
the yield of multiplex PCR (Fig.1) and multiplex HDA (Fig.2)
Improve
the processivity of DNA polymerase (10)
St
ABI
lization
and marking of ssDNA structure (11)
Increase
the yield and specificity of PCR reactions (5-9)
Increase
the yield and processivity of RT during RT-PCR (3,4)
Improve
DNA sequencing through regions with strong secondary structure (8)
Enhance
the RecA activity for ssDNA binding and strand trasfer (12,13)
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Storage Conditions:
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Concentration:
0.5mg/ml
Storage Buffer:
20 mM Tris-HCl
200 mM NaCl
1 mM EDTA
0.5 mM DTT
50% Glycerol
pH7.5 @ RT
Storage Temperature:
-20?C
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Quality Control:
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Quality Assurance Statement:
ET SSB is purified free of contaminating endonucleases and exonucleases.
Each lot is tested for single-strand, DNA-dependent ATPase activity
and is visually determined to be > 95% pure on an SDS-polyacrylamide
gel.
Exonuclease Activity:
Incubation of 20 ?g ET SSB for 4 hours at 37°C in 50 ?l
reaction buffer containing 50 mM potassium acetate, 20 mM Tris-acetate,
10 mM magnesium acetate and 1 mM dithiothreitol, pH 7.9 @ 25°C,
with 1 ?g of a mixture of single and double-stranded [
3
H]
E. coli
DNA (200,000 cpm/?g) released < 0.05% of
the total r
ADI
oactivity.
Endonuclease Assay:
Incubation of 10 ?g ET SSB for 4 hours at 37°C in 50 ?l
reaction buffer containing 50 mM potassium acetate, 20 mM Tris-acetate,
10 mM magnesium acetate and 1 mM dithiothreitol, pH 7.9 @ 25°C,
with 1 ?g ΦX174 RF I DNA gave < 5% conversion to RF
II.
Nuclease Activity:
Incubation of 20 ?g ET SSB for 16 hours at 37°C in 50 ?l
of reaction buffer containing 50 mM potassium acetate, 20 mM Tris-acetate,
10 mM magnesium acetate and 1 mM dithiothreitol, pH 7.9 @ 25°C,
with 1 ?g λ DNA yielded a clear and sharp band on an
agarose gel.
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References:
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Coleman, J. E. & Oakley, J. L. (1980) CRC Crit Rev Biochem
7, 247-89.
Chase, J. W. & Williams, K. R. (1986) Annu Rev Biochem 55,
103-36.
Baugh, L. R., Hill, A. A., Brown, E. L. & Hunter, C. P.
(2001) Nucleic Acids Res 29, E29.
Villalva, C., Touriol, C., Seurat, P., Trempat, P., Delsol,
G. & Brousset, P. (2001) Biotechniques 31, 81-3, 86.
Schwarz, K., Hansen-Hagge, T. & Bartram, C. (1990) Nucleic
Acids Res 18, 1079.
Chou, Q. (1992) Nucleic Acids Res 20, 4371.
Oshima, R. G. (1992) Biotechniques 13, 188.
Rapley, R. (1994) Mol Biotechnol 2, 295-8.
Olszewski, M., Rebala, K., Szczerkowska, Z. & Kur, J. (2005)
Mol Cell Probes 19, 203-5.
Myers, T. W. & Romano, L. J. (1988) J Biol Chem 263, 17006-15.
Delius, H., Mantell, N. J. & Alberts, B. (1972) J Mol Biol
67, 341-50.
Reddy, M. S., Vaze, M. B., Madhusudan, K. & Muniyappa, K.
(2000) Biochemistry 39, 14250-62.
West, S. C., Cassuto, E. & Howard-Flanders, P. (1982) Mol
Gen Genet 186, 333-8.
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產(chǎn)品貨號：1490.4
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