Reverse Transcription
Hifair™ Ⅲ 1st Strand cDNA Synthesis SuperMix for qPCR (gDNA Digester Plus) - 11141ES
A readjust premix developed based on Hifair™ⅢReverse Transcriptase. Compared with Hifair™ ⅡReverse Transcriptase, Hifair™ Ⅲ Reverse Transcriptase has significantly higher thermal stability and can withstand reaction temperatures up to 60℃, and is suitable for Reverse transcription of RNA templates with complex secondary structures. At the same time, the enzyme enhances the affinity with templates, which is ideal for the reverse transcription of a small number of templates and low-copy genes.
The premix contains 5×gDNA Digester Mix and 4×Hifair™ Ⅲ SuperMix Plus. 5×gDNA Digester Mix can remove residual genomic DNA contamination in RNA templates to ensure more reliable follow-up results. 4×Hifair™ Ⅲ SuperMix Plus contains all components required for Reverse Transcriptase (Buffer, dNTP, Hifair™ Ⅲ Reverse Transcriptase, RNase inhibitor, Random Primers/Oligo (dT)18 Primer Mix), RNA template and Rnase-free H2O are added to reverse transcription, and gDNA digester is terminated simultaneously to ensure the integrity of cDNA.
The product is suitable for two-step RT-QPCR detection, specially optimized for qPCR, proportionally optimized Random Primers/Oligo (dT)18 Primer Mix, enabling cDNA synthesis to be initiated from all regions of RNA transcript with the same reverse transcription efficiency. The authenticity and repeatability of qPCR results are guaranteed to the greatest extent. The reverse transcription product is compatible with SYBR Green method and TaqMan probe method qPCR, Hifair™ UNICON™ qPCR SYBR Green Master Mix, Hifair™ qPCR SYBR Green Master Mix or Hifair™ qPCR TaqMan Probe Master Mix can be selected according to experimental purposes, is used in combination to perform high-performance gene expression analysis. Figure 1. Excellent reverse transcription efficiency, suitable for genes with different expression abundances with different GC contents Figure 2. Linear assay range Total RNA 10 pg~5 μg Figure 3. Efficient removal of genomic residues Selected citations: [1] An RNA-cleaving threose nucleic acid enzyme capable of single point mutation discrimination. Nat Chem . 2022 Mar;14(3):350-359. doi: 10.1038/s41557-021-00847-3 [2] The tetratricopeptide repeat protein OsTPR075 promotes heading by regulating florigen transport in rice. Plant Cell . 2022 Sep 27;34(10):3632-3646. doi: 10.1093/plcell/koac190
The premix contains 5×gDNA Digester Mix and 4×Hifair™ Ⅲ SuperMix Plus. 5×gDNA Digester Mix can remove residual genomic DNA contamination in RNA templates to ensure more reliable follow-up results. 4×Hifair™ Ⅲ SuperMix Plus contains all components required for Reverse Transcriptase (Buffer, dNTP, Hifair™ Ⅲ Reverse Transcriptase, RNase inhibitor, Random Primers/Oligo (dT)18 Primer Mix), RNA template and Rnase-free H2O are added to reverse transcription, and gDNA digester is terminated simultaneously to ensure the integrity of cDNA.
The product is suitable for two-step RT-QPCR detection, specially optimized for qPCR, proportionally optimized Random Primers/Oligo (dT)18 Primer Mix, enabling cDNA synthesis to be initiated from all regions of RNA transcript with the same reverse transcription efficiency. The authenticity and repeatability of qPCR results are guaranteed to the greatest extent. The reverse transcription product is compatible with SYBR Green method and TaqMan probe method qPCR, Hifair™ UNICON™ qPCR SYBR Green Master Mix, Hifair™ qPCR SYBR Green Master Mix or Hifair™ qPCR TaqMan Probe Master Mix can be selected according to experimental purposes, is used in combination to perform high-performance gene expression analysis. Figure 1. Excellent reverse transcription efficiency, suitable for genes with different expression abundances with different GC contents Figure 2. Linear assay range Total RNA 10 pg~5 μg Figure 3. Efficient removal of genomic residues Selected citations: [1] An RNA-cleaving threose nucleic acid enzyme capable of single point mutation discrimination. Nat Chem . 2022 Mar;14(3):350-359. doi: 10.1038/s41557-021-00847-3 [2] The tetratricopeptide repeat protein OsTPR075 promotes heading by regulating florigen transport in rice. Plant Cell . 2022 Sep 27;34(10):3632-3646. doi: 10.1093/plcell/koac190
Features: Applications:
- High-performance reverse transcription
- Increase reaction temperature - First strand cDNA can be synthesized in the temperature range of 55 to 60°C
- Efficient removal of genomic residues
- Broad linear detection range
- Construction of cDNA library
- Antisense RNA synthesis
Murine RNase inhibitor (40 U/μL) - 10603ES
Murine RNase Inhibitor is purified from a recombinant strain of E. coli in a soluble form. It specifically inhibits the activity of RNases A, B and C through binding noncovalently in a 1:1 ratio with high affinity. Recombinant murine RNase inhibitor does NOT contain 2 oxidation-sensitive cysteine which are contained in human-origin RNase inhibitor. Therefore, murine RNase inhibitor has high anti-oxidation activity and is more stable for low DTT experiments (< 1mmol/L). This product is validated for its compatibility with HifairTM Ⅱ Reverse Transcriptase (Cat#11110) and various DNA Polymerases. Murine RNase inhibitor is ideal for high-DTT-sensitive experiments, such as RT-PCR. The product can be used in cDNA Synthesis, polysome isolation and In vitro transcription/translation.
Figure 1. RNase inhibitor can effectively inhibit RNase A activity to prevent the digestion of total RNA from HEK 293 cells.
Table 1. Examination of the inhibition of MRI on RNase A. Different concentration gradients of MRI were incubated with 100 ng of RNase A to block the activity of RNase A, followed by digestion with 1 μg of RNA and examined by one step RT-qPCR.
Selected citations:
[1] A fluorinated peptide with high serum- and lipid-tolerence for the delivery of siRNA drugs to treat obesity and metabolic dysfunction. Biomaterials. 2022;285:121541. doi:10.1016/j.biomaterials.2022.121541(IF:12.479)
[2] RBM39 Alters Phosphorylation of c-Jun and Binds to Viral RNA to Promote PRRSV Proliferation. Front Immunol. 2021;12:664417. Published 2021 May 17. doi:10.3389/fimmu.2021.664417(IF:7.561)
Features:
- Performs under a wide range of reaction conditions
- Protects RNA from degradation at temperatures up to 55°C
Hifair™ V Reverse Transcriptase - 11300ES
This is an updated vesion of HieffTM M-MLV (H-) Reverse Transcriptase obtained by genetic engineering technology. It has higher cDNA synthesis ability, thermal stability and reaction temperature limit (up to 60°C) than HieffTM M-MLV (H-) Reverse Transcriptase. The synthesized cDNA product is up to 10 kb. HifairTM V Reverse Transcriptase enhances the affinity of the templates and is suitable for reverse transcription of RNA templates with complex secondary structure or low copy genes.
Features: Applications:
- 5'→3' DNA polymerase activity
- 5'→3' exonuclease activity
- Hieff™ Taq DNA polymerase is an ideal tool for standard PCR with templates of 5 kb or less
- Genotyping
- Colont PCR
- Other conventional PCR
Hifair™ AdvanceFast 1st Strand cDNA Synthesis Kit (No Dye) - 11150ES
A rapid reverse transcription kit developed based on the Hifair™ III 1st Strand cDNA Synthesis Kit (No Dye), suitable for PCR amplification and RT-qPCR experiments. Compared to the Hifair™ III 1st Strand cDNA Synthesis Kit (No Dye), it boasts stable detection rates, specificity, and yield assurance, with the total reverse transcription time potentially reduced to less than 6 minutes, significantly shortening experimental duration.
The kit includes gDNA Digester Mix, which can eliminate genomic DNA contamination remaining in the RNA template, ensuring more reliable subsequent results. The kit provides two types of cDNA synthesis primers: Random Primers N6 and Oligo (dT)18. Users can choose between Random Primers N6, Oligo (dT)18, or Gene Specific Primers as reverse transcription primers according to their needs. The synthesized single-stranded cDNA product can be directly used for subsequent PCR or qPCR reactions.
The kit includes gDNA Digester Mix, which can eliminate genomic DNA contamination remaining in the RNA template, ensuring more reliable subsequent results. The kit provides two types of cDNA synthesis primers: Random Primers N6 and Oligo (dT)18. Users can choose between Random Primers N6, Oligo (dT)18, or Gene Specific Primers as reverse transcription primers according to their needs. The synthesized single-stranded cDNA product can be directly used for subsequent PCR or qPCR reactions.
Hifair™ AdvanceFast One-step RT-gDNA Digestion SuperMix for qPCR - 11151ES
This is an upgraded version of the Hifair™ V one-step RT-gDNA digestion SuperMix for qPCR, which meets the requirements of both reverse transcription and genomic DNA removal reactions in the same tube. The Hifair™ V one-step RT-gDNA digestion SuperMix for qPCR is an upgraded version of the Hifair™ V one-step RT-gDNA digestion SuperMix for qPCR, which allows for both reverse transcription and genomic DNA removal in the same tube, while simplifying the reaction process, dramatically shortening the reaction time, and increasing the yield of reverse transcription products. The 4×Hifair™ AdvanceFast One-Step RT SuperMix contains all the reagents required for the reverse transcription reaction, and only requires the addition of gDNA Remover Mix, template RNA, and water to efficiently synthesize the first-strand cDNA while removing the contamination of genomic DNA. It is suitable for reverse transcription of complex RNA templates as well as small amounts of template and low-copy genes.
This product is compatible with probe and dye-based qPCR. Hieff Unicon™ Universal TaqMan multiplex qPCR master mix (Cat#11211) and Hieff UNICON™ Universal Blue qPCR SYBR Green Master Mix (Cat#11184) are recommended for high-performance gene expression analysis. Hieff Unicon™ Universal TaqMan multiplex qPCR master mix (Cat#11211) and Hieff UNICON™ Universal Blue SYBR Green Master Mix (Cat#11184) for high performance gene expression analysis. The reverse transcription product is not suitable for subsequent gene cloning and other long fragment amplification, if needed, we recommend Hifair™ AdvanceFast 1st Strand cDNA Synthesis Kit (Cat#11149, Cat#11150) for high efficiency reverse transcription.
This product is compatible with probe and dye-based qPCR. Hieff Unicon™ Universal TaqMan multiplex qPCR master mix (Cat#11211) and Hieff UNICON™ Universal Blue qPCR SYBR Green Master Mix (Cat#11184) are recommended for high-performance gene expression analysis. Hieff Unicon™ Universal TaqMan multiplex qPCR master mix (Cat#11211) and Hieff UNICON™ Universal Blue SYBR Green Master Mix (Cat#11184) for high performance gene expression analysis. The reverse transcription product is not suitable for subsequent gene cloning and other long fragment amplification, if needed, we recommend Hifair™ AdvanceFast 1st Strand cDNA Synthesis Kit (Cat#11149, Cat#11150) for high efficiency reverse transcription.
Recombinant Deoxyribonuclease I (DNase I, RNase-free) - 10325ES
DNase I is an endonuclease that can digest single- or double-stranded DNA. It can hydrolyze phosphodiester bonds to produce mono- and oligodeoxynucleotides containing a 5'-phosphate group and a 3'-OH group.The optimal working pH range of DNase I is 7-8. The activity of DNase I depends on Ca2+ and can be activated by divalent metal ions such as Co2+, Mn2+, Zn2+, etc. In the presence of Mg2+, DNase I can randomly cleave any site of double-stranded DNA; while in the presence of Mn2+, DNase I can cleave DNA double-stranded at the same site, forming blunt ends or sticky ends with 1-2 nucleotides protruding. It can be used for the processing of various RNA samples.
Applications: QC:
- Using the human genome as a template, a fragment with a GC content of 72% was amplified using Taq DNA Polymerase.
- Strictly to ensure no detection of exonuclease or endonuclease residue
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