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.
Description
The DM1100 ExcelBand™ 50 bp DNA Ladder is a ready-to-use DNA ladder, which is pre-mixed with loading dye for direct gel loading. The DNA ladder DM1100 is composed of 17 individual DNA fragments: 1.5k, 1.2k, 1k, 900, 800, 700, 600, 500, 450, 400, 350, 300, 250, 200, 150, 100, and 50 bp derived from a mixture of PCR products and specifically digested plasmid DNA. This product contains two enhanced bands (500 bp and 200 bp) for easier reference. In addition, the low range Orange G tracking dye mimics the migration of a 50 bp dsDNA during electrophoresis is also added for real time monitoring.
Features
Source
Phenol extracted PCR products and dsDNA digested with specific restriction enzymes, equilibrated in 10 mM Tris-HCl (pH 8.0) and 10 mM EDTA.
Range
50 ~ 1,500 bp
Concentration
54 µg/ 500 µl
Recommended loading volume
5 µl/ well
Storage
Room temperature for 6 months
4°C for 12 months
-20°C for 36 months
The DM1100 ExcelBand™ 50 bp DNA Ladder is a ready-to-use DNA ladder, which is pre-mixed with loading dye for direct gel loading. The DNA ladder DM1100 is composed of 17 individual DNA fragments: 1.5k, 1.2k, 1k, 900, 800, 700, 600, 500, 450, 400, 350, 300, 250, 200, 150, 100, and 50 bp derived from a mixture of PCR products and specifically digested plasmid DNA. This product contains two enhanced bands (500 bp and 200 bp) for easier reference. In addition, the low range Orange G tracking dye mimics the migration of a 50 bp dsDNA during electrophoresis is also added for real time monitoring.
【Material】The vial storage box is made of durable cardboard with waterproof coating and cardboard dividers；chemically resistant to alcohols and mild organic
【Temperature Range】These freezer boxes are stable from -196°C to 121°C
【Applications】The freezer box is suitable for iquid nitrogen freezing (vapor phase)
【Capacity】Size: Height of 2 inch, Holds 0.5ml, 1.5ml, 2.0ml tubes
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