NZY Reverse Transcriptase
Opis
NZY Reverse Transcriptase is a modified recombinant form of the Moloney Murine Leukemia Virus (M-MuLV) Reverse Transcriptase purified from Escherichia coli. The enzyme has been modified in order to promote stability. NZY Reverse Transcriptase synthesizes the complementary DNA strand in the presence of a primer using either RNA (cDNA synthesis) or single-stranded DNA as a template at temperatures up to 50 °C. This is useful when using templates with high GC-content or with high degree of secondary structure. The enzyme lacks 3´→5´ exonuclease activity and has no RNase H activity, enabling improved synthesis of full-length cDNA even for long mRNA, using random priming.
Features
– Synthesizes of cDNA from RNA or ssDNA in only 30 min
– High yields of first-strand cDNA
– No intrinsic RNase H activity
– Thermostable – working temperature range 50-55 ºC
Applications
– First-strand cDNA synthesis for RT-PCR and RT-qPCR
– Reverse transcription at elevated temperatures (ideal for high GC-content templates)
– Synthesis of cDNA for cloning and expression
– Analysis of RNA by primer extension
High sensitivity of NZY Reverse Transcriptase:
The high sensitivity of NZY Reverse Transcriptase is demonstrated using decreasing amounts of total RNA isolated from mouse liver (1 μg – 10 pg) as starting material in a 20 μL first-strand cDNA synthesis reaction with 200 units of the enzyme.
After the reverse transcription, 1 μL of the resultant cDNA was used as a template to amplify the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene in a 25 μL end-point PCR reaction with Supreme NZYTaq DNA polymerase. NC: No template control. Lane M: NZYDNA Ladder III
Specifications
Product length: up to 7 kb
Sensitivity: high
Optimal reaction temperature: 50 ºC
Available as Kits: NZY First-Strand cDNA Synthesis Kit (MB125); NZY First-Strand cDNA Synthesis Kit, separate oligos (MB170); NZY First-Strand cDNA Synthesis Kit, no oligos (MB171)
Speed: 30 min
Denaturing conditions: NZY Reverse Transcriptase is inhibited in the presence of metal chelators (e.g. EDTA), inorganic phosphate, pyrophosphate and polyamines. The enzyme is inactivated at 85 °C for 5 min
Components
– NZY Reverse Transcriptase (200 U/μL)
– Reaction Buffer (10x)
NZYTech Reverse Transcriptases: Selection Guide
Dane techniczne
Opakowanie | 20 000 U |