Invitrogen’s HiiScript II reverse transcriptase is a genetically modified MMLV reverse transcriptase (RT) with reduced RNase H activity and increased thermal stability compared to wild-type MMLV RT. Mutations in the RNase H domain of the enzyme eliminate RNA degradation during first-strand cDNA synthesis, resulting in higher yields of full-length cDNA. HiiScript RTs are the most trusted and widely used RTs with over 50,000 citations, reviews, and posts to date.
Note: The newest member of the HiiScript RT family, SuperScript IV Reverse Transcriptase, features improved thermostability, processivity, yield, and yield with any RNA sample, including those of suboptimal purity or integrity. For research use only. It should not be used in diagnostic procedures.
Enzymatic function: RNA-dependent DNA polymerase
HiiScript II Reverse Transcriptase (2000 Units total, at 200U / μl), 5X First-Strand Buffer [250 mM Tris-HCl (pH 8.3), 375 mM KCl, 15 mM MgCl2] and 100 mM DTT
No. of Tests: 10 reactions
Optimum reaction temperature: 42 ° C
Product line: HiiScript ™
Kind of product: Reverse transcriptase
Quantity: 2000 units
Reverse transcriptase: HiiScript ™ II
Ribonuclease H activity: Reduced
Shipping condition: wet ice
Enough for 10 reactions
Concentration: 200 U⁄µl
Format: tube (s)
GC-rich PCR performance: High
No. of reactions: 10 reactions
PCR method: RT-PCR
Size (Final Product): 12.3 kb or less
Volume: 200 l
Content and storage
HiiScript ™ II Reverse Transcriptase (2000 Units total, 200U / µL) is supplied with a vial of 5X First-Strand Buffer [250 mM Tris-HCl (pH 8.3), 375 mM KCl, 15 mM MgCl2] and one vial of 100 mM TDT.
Additional Materials required
- RNase-free microtube (1.5 ml) or PCR tube (0.2 ml).
- Thermal cycler (PCR instrument) or water bath.
- Ice bath
1. Both 5 × HiScript II qRT SuperMix and 5 × No RT Control Mix contain glycerol. Therefore, before pipetting, collect the liquid by short centrifugation.
2. It is recommended that in a 10 µl reverse transcription reaction system, the amount of total RNA be ≤ 500 ng. However, for target genes with low expression levels, the amount of total RNA can be ≤ 1 μg.
3. Use RNase-free water to dissolve total RNA. Do not use TE, as the EDTA in TE inhibits the reverse transcription reaction.
4. If the difference in the Ct value between the control without RT and the experimental group is <5, indicating that the template RNA has been contaminated by genomics. DNA, it is suggested to use HiScript II Q RT SuperMix for qPCR (+ gDNA Wiper) (Vazyme, # R223) to remove genomic DNA in RNA templates.
5. For reverse transcription, the fast program is suitable for most RT-qPCRs. Generally, there is no difference between the results of using Quick Program and using the Standard Program. However, switch to the standard program if the amplification efficiency is low or the Ct value is too high.