The Opentrons® 3-piece Aluminum Block Set is used with our Temperature Deck. The set comes with a 24-well, 96-well and flat bottom plate format block. These are used to keep your 2mL tubes, 1.5mL tubes, 96-well PCR plates, PCR strips and flat bottom plates at a constant temperature. These blocks holds temperatures between 4°C and 95°C when used with the Opentrons Temperature Module (GEN2).
The Opentrons® 3-piece Aluminum Block Set is used with our Temperature Deck. The set comes with a 24-well, 96-well and flat bottom plate format block. These are used to keep your 2mL tubes, 1.5mL tubes, 96-well PCR plates, PCR strips and flat bottom plates at a constant temperature. These blocks holds temperatures between 4°C and 95°C when used with the Opentrons Temperature Module (GEN2).
Norgen’s microScript microRNA cDNA Synthesis Kit is an all-in-one, ready-to-use product for the reverse transcription of microRNA from either Total RNA preparations or enriched microRNA preparations. The kit contains the 2x Reaction Mix and the microScript microRNA Enzyme Mix. The kit utilizes Norgen’s microScript Reverse Transcriptase, a mutant version of Moloney Murine Leukemia Virus (M-MuLV) Reverse Transcriptase. It has reduced RNase H activity and increased thermal stability.
The workflow of Norgen’s microScript microRNA cDNA Synthesis Kit involves a simple, single-tube set-up by the mixing of 2x Reaction Mix, Enzyme Mix and the RNA template. The reaction can then be carried out in a thermocycler. A poly (A) tail is first added to the RNA template, followed by cDNA synthesis using an adapter primer. In addition to the ease-of-use, the single-tube set-up provides superb consistency and sensitivity. The cDNA could be used in a PCR or qPCR amplification using a Universal PCR Reverse Primer and the forward primer that contains the sequence of the microRNA of interest. A single cDNA preparation could be used for PCR amplification of a number of different microRNAs. In addition, the cDNA preparation could be used for PCR or qPCR detection (using gene-specific forward and reverse primers) of mRNA or large RNA if total RNA preparation was the starting template. This could allow for parallel evaluation of expression level of microRNAs and microRNA-targets.
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Storage Conditions and Product Stability
Norgen’s microScript microRNA cDNA Synthesis Kit components should be stored at -20°C. These reagents should remain stable for at least 1 year in their unopened containers.
| Component | Cat. 54415 (12 rxns) | Cat. 54410 (50 rxns) |
|---|---|---|
| microScript microRNA Enzyme Mix | 12 μL | 50 μL |
| 2x Reaction Mix | 120 μL | 500 μL |
| Universal PCR Reverse Primer | 60 μL | 250 μL |
| Nuclease-Free Water | 1.25 mL | 2 x 1.25 mL |
| Product Insert | 1 | 1 |
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.
PACE 2.0 Genotyping Master Mix ensures an unrivalled signal-to-noise ratio and produces tight data clusters, even when working with high-throughput, crude DNA preps, resulting in consistently exceptional performance. Efficiently streamline your workflow and reduce costs without compromising the quality of your results.
PACE 2.0 Genotyping Master Mix is an ideal solution for challenging starting material. PACE 2.0 has been specially formulated to overcome the obstacles presented by common PCR inhibitor compounds, such as phenols and tannins. Even notoriously tricky samples like oil palm and conifers can still be assayed using hot shot or other crude DNA prep methods and deliver reliable and accurate data.
PACE 2.0 Genotyping Master Mix uses a novel, universal, fluorescent reporting cassette to produce machine-readable fluorescent signals corresponding to genotypes. PACE 2.0 compatible genotyping assays are comprised of two competitive allele-specific forward primers (which differ in their terminal 3’ bases and unique 5’ tail sequences) and a common, reverse primer. PACE 2.0 Genotyping Master Mix is supplied with ROX normalising dye at a range of levels to ensure compatibility with your qPCR instrument or reader.
Genotyping assay designs are available from 3CR Bioscience through our free assay-design service; once designed, users can purchase assay primers independently or through 3CR Bioscience using our partial or full-assay validation service. PACE 2.0 Genotyping Master Mix is also compatible with KASP™ and Amplifluor® marker assays.
For Research and Development purposes only. Not for diagnostic use.
Legal Information
KASP™ is a trademark of LGC Biosearch Technologies
Amplifluor® is a registered trademark of Merck KGaA
