La transcriptase inverse M-MLV est un ADN polymérase recombinant qui synthétise un brin d’ADN complémentaire à partir d’ARN premier brin,Afficher plus
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Nbre de réactions
28025021
1 000 réactions
28025013
200 réactions
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Référence 28025021
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1 968,00
Each
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La transcriptase inverse M-MLV est un ADN polymérase recombinant qui synthétise un brin d’ADN complémentaire à partir d’ARN premier brin, d’ADN ou d’ARN / ADN hybride. Comparée à l’AMV RT, la transcriptase inverse du virus de la leucémie murine de Moloney (RT M-MLV) est dépourvue de toute activité de l’endonucléase d’ADN et affiche une activité RNase H inférieure. Fonctions de cette enzyme :
• Thermostabilité : activité optimale à 37°C • Taille de l’ADNc : M-MLV peut être utilisée pour synthétiser l’ADNc premier brin jusqu’à 7 kb • Applications : synthèse d’ADNc premier brin, extension de l’amorce, séquençage de l’ADNc, bibliothèques ADNc et RT-PCR
Source purifié à partir d’E. coli exprimant le gène pol de M-MLV sur un plasmide
Tests de performance et de qualité pureté SDS-PAGE ; tests d’endodésoxyribonucléase, d’exodésoxyribonucléase et de ribonucléase ; et rendement et longueur du produit ADNc
Définition d’unité une unité de RT M-MLV représente la quantité d’enzyme requise pour incorporer 1 nmol de désoxyribonucléotide dans une substance précipitable à l’acide, en 10 minutes, à 37°C, en utilisant poly(A) oligo(dT)25 comme modèle/amorce.
Conditions de réaction des unités 50 mM de Tris-HCl (pH 8,3), 40 mM de KCl, 6 mM de MgCl2, 1 mM de DTT, 0,5 mM de [3H]dTTP, 0,1 mM de poly(A), 0,1 mM d’oligo(dT)25, 0,1 mg/ml de BSA et enzyme dans 50 µl pendant 10 min à 37°C.
Usage exclusivement réservé à la recherche. Ne pas utiliser pour des procédures de diagnostic.
Spécifications
Type de produit finalADNc premier brin
FormatTube
Nbre de réactions1 000 réactions
Température de réaction optimale37°C
Quantité1000 réactions
Format de réactionComposants séparés
Type de réactifTranscription inverse
Transcriptase inverseM-MLV
Activité de la ribonucléase HOui
Conditions d’expéditionGlace humide
Taille (produit final)Jusqu’à 7 kb
Matériau de départARN
TechniqueTranscription inverse
Concentration200 U / μl
Vitesse de réaction50 min
Unit SizeEach
Contenu et stockage
• M-mLV RT, 1 ml (200 U/μl) • Tampon premier brin, 4 x 1 ml • DTT, 4 x 500 μl (100 mM)
Conserver à –20°C.
Foire aux questions (FAQ)
How much of the first-strand cDNA reaction should I load for PCR?
While the volume is dependent on the starting amount of RNA used for the first-strand synthesis and the abundance of the target gene, we'd recommend starting with 10% of the first-strand reaction for your PCR reaction.
What is the highest temperature that MMLV, SuperScript II, SuperScript III, or SuperScript IV RTs can be used?
The optimal temperature for for MMLV is 42 degrees C. The optimal temperature for SuperScript II RT is 42 degrees C, and can be used up to 50 degrees C. The optimal temperature for SuperScript III RT is 50 degrees C, and can be used up to 55 degrees C. For some qRT-PCR reactions where gene-specific primers are used, you can do the RT reaction at 60 degrees C. The optimal temperature for SuperScript IV RT is 50 degrees C, but can be used up to 65 degrees C.
Can I use a DNA-RNA hybrid as a template for M-MLV Reverse Transcriptase (Cat. No. 28025013, 28025021)? Can other reverse transcriptases, such as SuperScript reverse transcriptase, be used in the same way?
Yes, you can use a DNA-RNA hybrid as a template for M-MLV Reverse Transcriptase.
We have not tested this for SuperScript reverse transcriptases, so we cannot guarantee it would also work with those products.
This article can be used as a reference for additional information.
Can I use elevated temperatures for reverse transcription with Moloney Murine Leukemia Virus (M-MLV) reverse transcriptase?
Moloney Murine Leukemia Virus (M-MLV) Reverse Transcriptase is not well-suited for elevated temperatures. Typically, M-MLV RT operates optimally at temperatures around 37 degrees C to 42 degrees C.
To perform reverse transcription at elevated temperatures (e.g., 50 degrees C or higher), consider using thermostable reverse transcriptases such as SuperScript IV or Maxima H Minus, which are engineered to perform efficiently at higher temperatures.
Citations et références (4)
Citations et références
Abstract
Raloxifene Upregulated Mesangial Cell MMP-2 Activity via ER-ß Through Transcriptional Regulation.
Authors:Fang M, Wu XC, Huang W,
Journal:Cell Biochem Biophys
PubMed ID:23471663
'Raloxifene, a second-generation selective estrogen receptor modulator, exerts estrogen-like effects in specific tissues. In this present study, we examined the effect of raloxifene on mesangial cell matrix metalloproteinase-2 (MMP-2) activity in streptozotocin-induced diabetic mice. Raloxifene increased the MMP-2 level in a dose-dependent and receptor-mediated manner. An antibody against estrogen receptor-ß ... More
Single cell rt-PCR identification of odorant receptors expressed by olfactory neurons.
Authors:Malnic B,
Journal:Methods Mol Biol
PubMed ID:23585038
'Mammals have between 400 and 1,300 functional odorant receptor (OR) genes in their genomes. Each olfactory sensory neuron in the nose expresses only one single type of OR out of this vast repertoire. The OR expressed by an olfactory sensory neuron determines its functional activity and wiring to the olfactory ... More
Fast-mode duplex qPCR for BCR-ABL1 molecular monitoring: innovation, automation, and harmonization.
Authors:Gerrard G, Mudge K, Foskett P, Stevens D, Alikian M, White HE, Cross NC, Apperley J, Foroni L,
Journal:Am J Hematol
PubMed ID:22566190
Reverse transcription quantitative polymerase chain reaction (RTqPCR)is currently the most sensitive tool available for the routine monitoring of disease level in patients undergoing treatment for BCRABL1 associated malignancies. Considerable effort has been invested at both the local and international levels to standardise the methodology and reporting criteria used to assess ... More
Transforming growth factor-ß is required for vasculogenic mimicry formation in glioma cell line U251MG.
Authors:Ling G, Wang S, Song Z, Sun X, Liu Y, Jiang X, Cai Y, Du M, Ke Y,
Journal:Cancer Biol Ther
PubMed ID:22104964
Both vasculogenic mimicry (VM) and transforming growth factor-ß (TGFß) are positively correlated with malignancy in glioma. Accordingly, we supposed that TGFß might be related with VM, and aimed to detect whether TGFß could influence VM formation in two glioma cell lines U251MG and SHG44, which were different in malignancy. We ... More