What is Maxima reverse transcriptase?
Thermo Scientific Maxima Reverse Transcriptase (RT) is a. novel reverse transcription enzyme that was developed by Thermo Scientific through in vitro evolution of M-MuLV RT. The enzyme possesses an RNA and DNA-dependent polymerase activity as well as RNase H activity.
Does RT PCR need gel electrophoresis?
All Answers (7) No need of gel electrophoresis if you decided to go with Q RT PCR. Based on CT value and delta delta CT method you should be able to see the difference in expression of your genes up on treatment .
Which enzyme is used for cDNA synthesis?
enzyme reverse transcriptase
In molecular biology, complementary DNA (cDNA) is synthesised from an RNA template in a reaction catalysed by the enzyme reverse transcriptase (RTase). cDNA synthesis is the first step in many molecular biology workflows, such as gene expression studies using real-time PCR.
What is the difference between PCR and gel electrophoresis?
Using gel electrophoresis to visualize the results of PCR The results of a PCR reaction are usually visualized (made visible) using gel electrophoresis. Gel electrophoresis is a technique in which fragments of DNA are pulled through a gel matrix by an electric current, and it separates DNA fragments according to size.
Why is oligo dT an effective primer for reverse transcriptase?
Oligo(dT) primers amplify only mRNAs containing a poly(A) tail, since that is where the primer binds to promote reverse transcription. Random primers amplify most RNA species, including degraded RNA and viral genomes.
What is M-MuLV reverse transcriptase?
Moloney Murine Leukemia Virus (M-MuLV, MMLV) Reverse Transcriptase is an RNA-directed DNA polymerase. This enzyme can synthesize a complementary DNA strand initiating from a primer using either RNA (cDNA synthesis) or single-stranded DNA as a template. (1-4). M-MuLV Reverse Transcriptase lacks 3´ → 5´ exonuclease activity.
What is the correct buffer for MMLV RT?
Provided with 5X Reaction Buffer: 250mM Tris-HCl (pH 8.3 at 25°C), 375mM KCl, 15mM MgCl 2, 50mM DTT. May Be Heat-Inactivated: M-MLV RT is inactivated by heating at 70°C for 10 minutes.
What is the correct buffer size for reverse transcriptase?
M-MLV Reverse Transcriptase Buffer is supplied as 5X Buffer [250 mM Tris-HCl (pH 8.3), 375 mM KCl, 15 mM MgCl2] with 500 µl 0.1 M DTT.
What is the RNase H activity of M-MLV RT?
The RNase H activity of M-MLV RT is weaker than that of the commonly used Avian Myeloblastosis Virus (AMV) reverse transcriptase. Provided with 5X Reaction Buffer: 250mM Tris-HCl (pH 8.3 at 25°C), 375mM KCl, 15mM MgCl 2, 50mM DTT.