Permagen’s 6 x 1.5 mL Microfuge Tube Magnetic rack is designed for magnetic bead separations from up to 6 tubes
Detail
Permagen’s 6 x 1.5 mL Microfuge Tube Magnetic rack is designed for magnetic bead separations from up to 6 tubes
Accommodates many common 1.5 mL Microcentrifuge and some 2.0 mL tubes
Tubes are angled and beads will be pulled to back wall allowing easy aspiration and tip tracking down the front wall of the tubes without disturbing bead pellet
Plasmid DNA is ready for various downstream applications including restriction digestion, bacterial transformation, sequencing and more
Available in 4 formats: MiniPrep, MiniPrep (Magnetic Bead System), 96-Well MiniPrep (Magnetic Bead System), and MaxiPrep
These kits are designed for the rapid preparation of plasmid DNA from Escherichia coli.
Plasmid MiniPrep Kit
This kit is designed for the rapid preparation of plasmid DNA from small cultures of Escherichia coli using convenient spin columns. The plasmid DNA is preferentially purified from other cellular components such as genomic DNA and RNA. This kit is able to purify plasmids up to 13,000 bp in size, and the typical purification yield is up to 20 μg from 1.5 mL of bacterial culture. Purified DNA is of excellent quality for transformation, restriction enzyme digestion, sequencing and more. Also available in a 96-well format.
Plasmid MiniPrep Kit (Magnetic Bead System and High Throughput Magnetic Bead System)
Norgen’s Plasmid MiniPrep Kit (Magnetic Bead System) is designed for the rapid preparation of plasmid DNA from small batch cultures of Escherichia coli. Norgen’s magnetic beads bind DNA under optimized salt concentrations and release the bound DNA under low salt and slightly alkali conditions. The plasmid DNA is preferentially purified from other cellular components such as genomic DNA and RNA. The purified plasmids are fully digestible with all restriction enzymes tested, and are completely compatible with real-time PCR and NGS.
Norgen’s Plasmid MiniPrep Kit (Magnetic Bead System) is also available in a 96-well (HT) format for high throughput applications. Purification with the 96-well plates can be integrated with a robotic automation system.
Plasmid MaxiPrep Kit
This kit is designed for the rapid spin column preparation of plasmid DNA from up to 100 mL of Escherichia coli cultures. The kit allows for the isolation of plasmid DNA with final endotoxin levels of 0.1 EU/µg of DNA or less. The kit is able to purify plasmids up to 13,000 bp in size, and typical yields from a 100 mL culture for a high copy number plasmid are between 0.4 and 1.0 mg. The purified DNA is fully digestible with all restriction enzymes tested, and is completely compatible with manual or automated sequencing to achieve 95-100% accuracy.
Storage Conditions and Product Stability All solutions should be kept tightly sealed and stored at room temperature. The RNase vial should be stored at -20°C upon arrival. Once RNase has been added to the Resuspension Solution AZ the solution should be stored at 4°C. This kit is stable for 1 year after its date of shipment.
Component
Cat. 13300 (50 preps)
Cat. 46400 (250 preps)
Cat. 46500 (4 preps)
Cat. 46600 (20 preps)
Cat. 60300 (50 preps)
Cat. 63000 (192 preps)
Resuspension Solution AZ
12 mL
60 mL
20 mL
100 mL
12 mL
2 x 20 mL
Lysis Buffer N
40 mL
80 mL
40 mL
2 x 80 mL
40 mL
2 x 40 mL
Buffer TN
20 mL
130 mL
55 mL
2 x 130 mL
20 mL
1 x 55 mL 1 x 20 mL
Wash Solution E
12 mL
2 x 18 mL
–
–
–
–
Elution Buffer K
8 mL
30 mL
–
–
8 mL
2 x 8 mL
Wash Solution J
–
–
25 mL
3 x 25 mL
–
–
Elution Buffer J
–
–
24 mL
120 mL
–
–
RNase A
1 vial
1 vial
1 vial
1 vial
1 vial
1 vial
Magnetic Bead Suspension
–
–
–
–
1 x 1.1 mL
4 x 1.1 mL
Spin Columns
50
250
–
–
–
–
Collection Tubes
50
250
–
–
–
–
DNA Maxi Spin Columns with Collection Tubes (Clear ring in column)
–
–
4
20
–
–
Maxi Spin Filter Columns with Collection Tubes (Grey ring in column)
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%.
Document
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.
