5X all-in-one RT MasterMix
From RNA template to cDNA in just 15 minutes, hassle-free. With ABM’s 5X All-In-One RT MasterMix, simply add RNA and this optimized system will provide sensitive and reliable cDNA synthesis in a dynamic range of input RNA. The use of this MasterMix eliminates the addition of multiple components providing exceptional reproducibility and precision, as well as offering the end-user maximum comfort.
Product description
5X all in one 1st Strand cDNA Synthesis Mix is an optimized MasterMix formulation including all the reagents necessary for the synthesis of the first-strand cDNA, with the exception of RNA. This 5X concentrated reaction master mix contains PowerScript ™ Reverse Transcriptase (RTase), RNaseOFF ribonuclease inhibitor, dNTP and an optimized ratio of Oligo (dT) s and random primers.
PowerScript RTase is a specially designed derivative of the native RTase enzyme of the Moloney murine leukemia virus, MMLV), which catalyzes the synthesis of complementary DNA strands from single-stranded RNA / DNA templates. RNAse H activity intrinsic to native RTase involves degradation of RNA during first-strand cDNA synthesis. Strategic Mutation Series made for PowerScript ™ RTase, including those for removal of RNase H activity, result in higher yields and an increase in the achievable length of synthesized cDNA. PowerScript ™ RTase also contains a fidelity enhancing subunit that ensures superior precision in reverse transcription.
The RNaseOFF ribonuclease inhibitor in the mix adds effective RNA template protection of any possible degradation by RNases. Regarding the primers, the Oligo (dT) s selectively hybridization to the poly (A) tail of mRNA; and the Random Primer, with its nonspecific character hybridization, allows the use of any type of RNA as a template.
Note:
Once the cDNA synthesis of the first strand is complete, the cDNA product can be applied directly as a template on a standard PCR / qPCR.
Storage conditions
Store all components at -20 ° C. All components are stable for 1 year from the date of shipping when properly stored and handled.
Primer information
Oligo (dT) are oligonucleotides that hybridize to the 3′-Poly (A) tail of mRNAs. The utility of the oligo (dT) is restricted to 3′-Poly (A) tailed mRNA or total RNA templates. Random Primers are collected at non-specific sites within the RNA template (s), can be used for all forms of RNA as a template for cDNA synthesis.
Key features
• Reduced handling errors with just 1 liquid transfer step
• Optimized protocol suitable for high-performance applications
• Simple configuration for any RNA template
• High reproducibility and excellent performance.
Package package options
• With the AccuRT Genomic DNA Removal Kit (Cat. # G492), remove contaminating genomic DNA from your samples in less than 10 minutes.
• With ExCellenCT Lysis Kit (Cat. No. G916) it allows extraction and preparation of RNA templates directly from cultured cells in less than 15 minutes, revealing RNA / reverse transcription-ready cell lysate from cultured cells without the dangerous and time-consuming chemicals -steps of extraction and purification of involved RNA.