Product Details
Application:
DNA Nucleic Acid
Format:
NFO Kit
Kit Components:
Reagents,Buffer,Enzymes
Manufacturer:
Amp-future Bio
Product Name:
DNA Isothermal Rapid Amplification Kit (Colloidal Gold Test Strip Type)
Reaction Time:
5-20mins
Reaction Volume:
50μL
Sensitivity:
500-1000copies/ml
Shelf Life:
14 Months
Specificity:
High
Storage Conditions:
-20℃
Suitability:
Universal
High Light:
,
,
Payment & Shipping Terms
Minimum Order Quantity
48T
Price
USD$3.8/T
Packaging Details
16T/Bag,48T/Box
Delivery Time
6 working days
Payment Terms
T/T, MoneyGram
Supply Ability
100000T/Month
Product Description
【Principle Overview】
This kit is based on a rapid nucleic acid amplification technology at room temperature and constant temperature: at room temperature and constant temperature (generally 39℃~42℃), with the help of auxiliary proteins and single-strand binding proteins, the recombinase and primers form a complex; Source search and combine the target homology domain, at this time, a D-loop region is formed at the homology position and strand exchange begins; along with the dissociation of the recombinase from the complex, the polymerase also binds to the 3′ end of the primer and begins chain extension. Relying on the action of nfo enzyme, adding specific molecular probes designed according to the template, and using colloidal gold technology (sandwich method) can detect the final result.
【Primer design】
It is recommended to use primers with a length of 30-35 bp.Too short primers will affect the amplification speed and detection sensitivity;the 5′ end of the reverse primer is labeled with a modification group (commonly used biotin).
Primers are designed to avoid the formation of secondary structures that affect amplification; the length of the amplicon is recommended to be 150-500bp.
【Colloidal gold probe design】
In the middle of the forward and reverse primer,design a sequence of 46-52nt in length that is complementary to the target fragment; modify an antigen marker (typical FAM) at the 5′ end;mark a dSpacer (tetrahydrofuran, THF) in the middle of the 5′ end and the 3′ end ), as the recognition site of nfo;the 3′ end is labeled with a modification group, such as amine group, phosphate group or C3-Spacer, etc.
【Features】
This kit has the advantages of high sensitivity, strong specificity,and short reaction time (only 15 minutes),and the reaction components are in dry powder state,which is easy to operate and easy to store.
This reagent has low equipment requirement,and the reaction operation can be carried out in metal baths, water baths, etc.,and there is no need to purchase expensive exclusive equipment such as PCR amplifiers.
| Composition | Content |
| A buffer | 1.6mL×1Tube |
| B buffer | 150μL×1Tube |
| Positive control template-II | 100μL×1Tube |
| Positive control probe and primer mix-II | 70μL×1Tube |
| Reagent | 48 T |
| Guide Manual | 1 Copy |
【Kit storage】
1. Storage conditions: storage temperature ≤ -20℃constant temperature environment, keep away from light and avoid heavy pressure;
2. Product validity period: 14 months;
3. See the outer packaging for the production date.
This kit is based on a rapid nucleic acid amplification technology at room temperature and constant temperature: at room temperature and constant temperature (generally 39℃~42℃), with the help of auxiliary proteins and single-strand binding proteins, the recombinase and primers form a complex; Source search and combine the target homology domain, at this time, a D-loop region is formed at the homology position and strand exchange begins; along with the dissociation of the recombinase from the complex, the polymerase also binds to the 3′ end of the primer and begins chain extension. Relying on the action of nfo enzyme, adding specific molecular probes designed according to the template, and using colloidal gold technology (sandwich method) can detect the final result.
Description
Q-PAGE™ TGN (Tris-Glycine Novel) Precast Gels are ready-to-use acrylamide gels for SDS-PAGE running in Tris-Glycine buffer system. With unique formula, Q-PAGE™ TGN Precast Gels perform enhanced speed, better separation, and longer shelf life as compared with conventional Laemmli Tris-HCl gels. The protein migration patterns in Q-PAGE™ TGN series, however, are similar with typical Laemmli Tris-HCl gels, and thus Q-PAGE™ TGN Precast Gels are compatible to traditional SDS-PAGE and subsequent analyses.
Q-PAGE™ TGN Precast Gels are available in gradient (4 to 15%) and fixed (10%) concentrations of polyacrylamide in 12- and 15-well formats. Two available cassette sizes, Mini (10 x 8.3 cm) and Midi (10 x 10 cm), are compatible with most popular protein electrophoresis systems. Q-PAGE™ Mini (QP4XXX) Gels are suitable for Bio-Rad® and other systems. Q-PAGE™ Midi (QP5XXX) Gels are suitable for Invitrogen® XCell SureLock® Mini-Cell, Invitrogen® Mini Gel Tank, Hoefer SE260, and other systems.
Key Features
Storage and stability
Store Q-PAGE™ Precast Gels at 4°C for periods up to 12 months.
Do not freeze Q-PAGE™ Precast Gels. Remove tape and comb before electrophoresis.
Keep Q-PAGE™ Precast Gels flat during storage.
Q-PAGE™ TGN (Tris-Glycine Novel) Precast Gels are ready-to-use acrylamide gels for SDS-PAGE running in Tris-Glycine buffer system. With unique formula, Q-PAGE™ TGN Precast Gels perform enhanced speed, better separation, and longer shelf life as compared with conventional Laemmli Tris-HCl gels. The protein migration patterns in Q-PAGE™ TGN series, however, are similar with typical Laemmli Tris-HCl gels, and thus Q-PAGE™ TGN Precast Gels are compatible to traditional SDS-PAGE and subsequent analyses.
Endonucleases Non-Specific, HL-SAN
HL-SAN efficiently removes nucleic acids from buffers typically used in protein purification. Due to its high salt tolerance, it is the obvious choice for host-cell DNA removal in settings where salt is added to reduce aggregation. Especially efficient for removing nucleic acids from proteins with high affinity for DNA and RNA. Proven performance during lysis and early stages of protein purification processes, as well as high-salt eluates. Cold-adapted enzyme with excellent performance also at ambient temperatures and during over-night digestion at 4°C.
Figure 1. Optimum activity in solutions with high salinity
HL-SAN has optimum activity at ∼0.5 M NaCl, but operates at a broad range of [NaCl] and [KCl]. The activity of HL-SAN was tested in a 25 mM Tris-HCl buffer, pH 8.5, 5 mM MgCl2 with varying [NaCl] or [KCl]. The maximum activity was set to 100%.
Figure 2. Temperature and activity
HL-SAN has optimum activity at ~35°C, but works over a broad temperature range (20% activity at 10°C and 50°C). The activity of HL-SAN was tested in a 25 mM Tris-HCl buffer, pH 8.5 containing 5 mM MgCl2 and 0.5 M NaCl.
Fig 3. The effect of MgCl2 and MnCl2 concentration on the HL-SAN activity.
The activity of HL-SAN was tested in a 25 mM Tris-HCl buffer, pH 8.5, 0.5 M NaCl and with varying concentrations of MgCl2 or MnCl2. The activity of the sample containing 5 mM MgCl2 was set to 100%.
Figure 4. HL-SAN activity vs pH/[NaCl]
The activity of HL-SAN was tested in a 25 mM Tris-HCl buffer with different pHs and different concentrations of NaCl. All buffers contained 5 mM MgCl2. The nature of the buffer was pH-dependent, but generally the NaCl-optimum was the same in all buffers/pHs. The exception was etanolaminbuffer at pH 9 and pH 9.5 in which the NaCl-optimum was shifted to the left (not shown).
Figure 5. Buffer composition affects substrate preference
Without NaCl, the specificity towards ssDNA and dsDNA is similar. At 0.5 M NaCl, the activity towards dsDNA increases, while the activity towards ssDNA is unaffected.
Figure 6. HL-SAN digests ssDNA to ~5-13 nt, and dsDNA to ~5-7 nt
The size of the end products from ssDNA varies from ~5-13 nt, while dsDNA is digested to around ~5-7 nt. The size of the end products seems to depend on the DNA sequence. Substrates 1 and 2 were ssDNA with different sequences and substrates 3 and 4 were dsDNA with similar sequences but with a FAM-label at different ends. Substrate 5 was dsDNA with the same sequence as substrate 3 and 4 but with a FAM-label at both ends.
Figure 7. HL-SAN activity decreases with increasing concentrations of glycerol
The activity of HL-SAN was tested in a 25 mM Tris-HCl buffer, pH 8.5, 5 mM MgCl2, 0.5 M NaCl and with increasing concentrations of glycerol. The activity of the control not containing glycerol was set to 100%.
Figure 8. The activity of HL-SAN at different concentrations of imidazole
The activity of HL-SAN was tested in a 25 mM Tris-HCl buffer, pH 8.5, 5 mM MgCl2, 0.5 M NaCl and with varying concentrations of imidazole. The activity of the control not containing imidazole was set to 100%.
HL-SAN efficiently removes nucleic acids from buffers typically used in protein purification. Due to its high salt tolerance, it is the obvious choice for host-cell DNA removal in settings where salt is added to reduce aggregation. Especially efficient for removing nucleic acids from proteins with high affinity for DNA and RNA. Proven performance during lysis and early stages of protein purification processes, as well as high-salt eluates. Cold-adapted enzyme with excellent performance also at ambient temperatures and during over-night digestion at 4°C.
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Tel : 081-875-1869 , 02-328-7179
Email : hej@a3p-scientific.com
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