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Citations & References (7)
Invitrogen™
SuperScript™ III First-Strand Synthesis System
The SuperScript III First-Strand Synthesis System for RT-PCR is optimized to synthesize first-strand cDNA from purified poly(A)+ or total RNA.Read more
| Catalog Number | Quantity |
|---|---|
| 18080051 | 50 rxns |
Catalog number 18080051
Price (EUR)
920,00
Each
Quantity:
50 rxns
Price (EUR)
920,00
Each
The SuperScript III First-Strand Synthesis System for RT-PCR is optimized to synthesize first-strand cDNA from purified poly(A)+ or total RNA. RNA targets from 100 bp to >12 kb can be detected with this system and the amount of starting material can vary from 1 pg to 5 μg of total RNA. SuperScript III Reverse Transcriptase is a version of M-MLV RT that has been engineered to reduce RNase H activity and provide increased thermal stability. The enzyme is used to synthesize cDNA at a temperature range of 42–55°C, providing increased specificity, higher yields of cDNA, and more full-length product than other reverse transcriptases. Because SuperScript III RT is not significantly inhibited by ribosomal and transfer RNA, it may be used to synthesize first-strand cDNA from a total RNA preparation.
Using the SuperScript III First-Strand System
cDNA synthesis is performed in the first step using either total RNA or poly(A)+-selected RNA primed with oligo(dT), random primers, or a gene-specific primer. In the second step, PCR is performed in a separate tube using primers specific for the gene of interest. For the PCR reaction, we recommend one of the following DNA polymerases: Platinum Taq DNA Polymerase provides automatic hot-start conditions for increased specificity up to 4 kb, Platinum Taq DNA Polymerase High Fidelity provides increased yield and high fidelity for targets up to 15 kb, and Platinum Pfx DNA Polymerase provides maximum fidelity for targets up to 12 kb.
For Research Use Only. Not for use in diagnostic procedures.
Specifications
Final Product TypeFirst-Strand cDNA
FormatKit
No. of Reactions50 Reactions
Optimal Reaction Temperature50°C
Quantity50 rxns
Reaction FormatSeparate components
Reagent TypeReverse Transcription
Reverse TranscriptaseSuperScript III
Shipping ConditionDry Ice
Starting MaterialRNA
TechniqueReverse Transcription
For Use With (Application)Real Time PCR (qPCR), RT-PCR
GC-Rich PCR PerformanceHigh
Reaction Speed50 min.
Unit SizeEach
Contents & Storage
• Oligo(dT)20, 50 μL (50 μM)
• Random hexamers, 250 μL (50 ng/μL)
• 10X RT buffer, 1 mL
• DTT, 250 μL (0.1 M)
• Magnesium chloride, 500 μL (25 mM)
• dNTP mix, 250 μL (10 mM)
• SuperScript III RT, 50 μL (200 U/μL)
• RNaseOUT, 100 μL (40 U/μL)
• E. coli RNase H, 50 μL (2 U/μL)
• DEPC-treated water, 1.2 mL
• Total HeLa RNA, 20 μL (10 ng/μL)
• Sense Control Primer, 25 μL (10 μM)
• Antisense Control Primer, 25 μL (10 μM)
Store at –20°C.
Frequently asked questions (FAQs)
I am interested in generating cDNA from total RNA. What is the difference between SuperScript III Reverse Transcriptase and SuperScript III First Strand Synthesis System for RT-PCR?
How long can I store the cDNA from my reverse transcription step?
How can I remove genomic DNA contamination from my sample prior to performing RT-PCR?
How much RNA should be employed for first-strand cDNA synthesis?
Should I treat the cDNA with RNase H prior to downstream processing?
Citations & References (7)
Citations & References
Abstract
The UL41 protein of herpes simplex virus 1 degrades RNA by endonucleolytic cleavage in absence of other cellular or viral proteins.
Journal:Proc Natl Acad Sci U S A
PubMed ID:16477041
The herpes simplex virus 1 ORF UL41 encodes a protein (virion host shutoff or vhs) associated with selective degradation of mRNA early in infection. Some mRNAs, exemplified by GAPDH or beta-actin mRNAs, are degraded rapidly. Others, for example IEX-1 mRNA, are degraded in two stages: whereas the 3' domain disappears
Transcriptional profiling of rhesus monkey embryonic stem cells.
Journal:Biol Reprod
PubMed ID:16943365
Embryonic stem cells (ESCs) may be able to cure or alleviate the symptoms of various degenerative diseases. However, unresolved issues regarding survival, functionality, and tumor formation mean a prudent approach should be adopted towards advancing ESCs into human clinical trials. The rhesus monkey provides an ideal model organism for developing
Novel GC-rich DNA-binding compound produced by a genetically engineered mutant of the mithramycin producer Streptomyces argillaceus exhibits improved transcriptional repressor activity: implications for cancer therapy.
Journal:Nucleic Acids Res
PubMed ID:16571899
The aureolic acid antibiotic mithramycin (MTM) binds selectively to GC-rich DNA sequences and blocks preferentially binding of proteins, like Sp1 transcription factors, to GC-rich elements in gene promoters. Genetic approaches can be applied to alter the MTM biosynthetic pathway in the producing microorganism and obtain new products with improved pharmacological
Temperature-modulated diversity of TRPV4 channel gating: activation by physical stresses and phorbol ester derivatives through protein kinase C-dependent and -independent pathways.
Journal:J Biol Chem
PubMed ID:12738791
The TRPV4 calcium-permeable channel was cloned from mouse kidney M-1 cells, and the effect of temperature modulation on channel gating/activation by physical and chemical signals was evaluated. A TRPV4 cDNA construct with a C-terminal V5 epitope was stably transfected into human embryonic kidney (HEK) 293 and Chinese hamster ovary cells
Endogenous 24(S),25-epoxycholesterol fine-tunes acute control of cellular cholesterol homeostasis.
Journal:J Biol Chem
PubMed ID:17981807
Certain oxysterols, when added to cultured cells, are potent regulators of cholesterol homeostasis, decreasing cholesterol synthesis and uptake and increasing cholesterol efflux. However, very little is known about whether or not endogenous oxysterol(s) plays a significant role in cholesterol homeostasis. 24(S),25-Epoxycholesterol (24,25EC) is unique among oxysterols in that it is