Heat-Labile Exonuclease I (HL-ExoI) is a 3’ – 5’ exonuclease, specific for single stranded DNA. The enzyme is recombinantly produced in E. coli. HL-ExoI is active at 25 – 37°C and inactivated by 1 minute incubation at 80°C or 15 minutes at 60°C.
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Heat-Labile Exonuclease I (HL-ExoI) is a 3’ – 5’ exonuclease, specific for single stranded DNA. The enzyme is recombinantly produced in E. coli. HL-ExoI is active at 25 – 37°C and inactivated by 1 minute incubation at 80°C or 15 minutes at 60°C.
HL-ExoI is used for degradation of ssDNA such as primers and oligos. It is also ideal for treatment of sensitive samples and useful in the development of novel molecular diagnostics applications.
Key Features
3’-5’ exonuclease specific for single stranded DNA
High activity at 25 – 37°C
Easily heat-inactivated by 1 min incubation at 80°C, or 15 min at 60°C
Moderate salt tolerance
Application
Removal of primers post-PCR prior to DNA sequencing or SNP detection
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Properties
Quality Control
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Endonucleases Non-Specific, HL-SAN
Product Info
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Product Info
Endonucleases Non-Specific, HL-SAN
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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.
Optimum activity at high salt concentration (0.5 M NaCl)
Active at low temperatures (20% at 6ºC)
Easily inactivated
Broad pH range
Temperature stable
Figures
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).
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%.
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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.
Extract high quality & quantity total RNA including miRNA
No phenol step required; isolate all RNA in one fraction
Bind & elute all RNA irrespective of size or GC content, without bias
Very sensitive & linear down to a few cells without the need for carrier RNA
Isolate from a wide variety of specimens
Purified RNA is suitable for a variety of downstream applications, including Small RNA Sequencing. Find out more information on Norgen’s NGS services
Available in a variety of formats to suit your needs
Purification is based on spin column chromatography that uses Norgen’s resin separation matrix
These kits are suitable for the isolation of total RNA from a range of samples including cells, bacteria, yeast, virus and bodily fluids including plasma/serum, blood, saliva, CSF and more. Extract high quality and purity RNA with excellent RIN values and A260/A280 suitable for downstream applications including qRT-PCR, RT-PCR, microarrays, NGS and more. These kits purify all sizes of RNA from large mRNA, lncRNA down to microRNA (miRNA) in the same fraction without the requirement of phenol. Isolate all RNA sequences at an equal rate irrespective of size. Moreover, when the RNA sequences are small (e.g. miRNA), the column binds small RNAs regardless of their GC content.
Total RNA Purification 96-Well Kit (High Throughput and High Throughput Deep Well)
This 96-well kit provides a rapid method for the high-throughput isolation and purification of total RNA in 30 minutes using vacuum manifold, plate centrifuge, or liquid handlers with vacuum capabilities. Total RNA can be isolated from a broad range of sample sources including cultured cells, tissues, blood, serum, plasma, bacteria, yeast, fungi, and viruses.
* for isolating total RNA from larger amounts of tissue, please use Norgen’s Animal Tissue RNA Purification Kit (Cat# 25700)
Storage Conditions and Product Stability All solutions should be kept tightly sealed and stored at room temperature. These kits are stable for 2 years after the date of shipment.
Q-PAGE™ Bis-Tris Precast Gel is a high-performance and easy to use precast polyacrylamide gel for electrophoresis in Bis-Tris buffer system (MOPS or MES). The optimized gel formula allows Q-PAGE™ Bis-Tris Precast Gel to show improved resolution, accurate results, and an extended shelf-life over conventional Laemmli Tris-HCl gels.
Q-PAGE™ Bis-Tris Precast Gels are available in gradient (4 to 12%) and fixed (8% and 12%) 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 (QP2XXX) Gels are suitable for Bio-Rad® and other systems. Q-PAGE™ Midi (QP3XXX) Gels are suitable for Invitrogen® XCell SureLock® Mini-Cell, Invitrogen® Mini Gel Tank, Hoefer SE260, and other systems.
Key Features
User-friendly gel cassette:
Numbered and framed wells for sample loading
With cassette opener for easy use
Enhanced gel performance:
Enhanced band sharpness
Better resolution of small proteins
Stable for shipping at ambient temperature
Easy compatibility:
Available as homogeneous and adjusted gradient gels for a wide range of protein separation.
Compatible with most popular protein electrophoresis systems
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.
Document
Q-PAGE™ Bis-Tris Precast Gel is a high-performance and easy to use precast polyacrylamide gel for electrophoresis in Bis-Tris buffer system (MOPS or MES). The optimized gel formula allows Q-PAGE™ Bis-Tris Precast Gel to show improved resolution, accurate results, and an extended shelf-life over conventional Laemmli Tris-HCl gels.
Q-PAGE™ Bis-Tris Precast Gels are available in gradient (4 to 12%) and fixed (8% and 12%) 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 (QP2XXX) Gels are suitable for Bio-Rad® and other systems. Q-PAGE™ Midi (QP3XXX) Gels are suitable for Invitrogen® XCell SureLock® Mini-Cell, Invitrogen® Mini Gel Tank, Hoefer SE260, and other systems.