dUTP solution 100mM

Oligo (dT)20 are single stranded oligodeoxyribonucleotides containing only deoxythymine (dT) to be able to prime with the poly(A) tail of mRNA molecules. Primers are quality controlled (MS), purified (reversed phase HPLC), aliquoted and lyophilized and came with a guaranteed amount of at least 5OD (about 27.2nmol (163.7µg)). Oligo (dT)20 Primer is designed to initiate the synthesis of cDNA from total RNA in a reverse transcription reaction, where Reverse Transcriptase, e.g. MMuLV (M3042) > is starting the reaction from the poly-A-end of mRNA. It can also be used for generation of labeled cDNA to screen microarrays. A mixture (1:1 ratio) of random hexamer primers (M3038 >) and Oligo(dT)20 primer may improve the sensitivity of cDNA synthesis as it will especially improve the efficiency of transcripts longer than 600 bp. Guaranteed delivery of at least 5OD (equals about 27.2nmol (163.7µg)). Primers are quality controlled (MS), purified (reversed phase HPLC), lyophilised and aliquoted. Dissolve content (5 OD) in 271.5μL of molecular biology grade water for an end concentration of 100 pmol/μL (100μM). 760μL (100μM) are sufficient for 760 reverse transcriptions of 20μL each. Oligo (dT)20 Primer is suitable for use as a primer for first strand cDNA synthesis with a reverse transcriptase, such as MMuLV or AMV. The primer hybridizes to the poly-adenylated tail found on the 3´ end of most eukaryotic mRNAs. Oligo (dT)20 ensures that the 3´ end of mRNAs are represented. Oligo(dT)20 Primer may be better suited for the new generation of Reverse Transcriptases like SuperScript™ that work at higher temperatures as the longer Oligo(dT)20 Primer enable annealing in reverse transcription reactions at higher temperatures. Applications• cDNA synthesis from total RNA in a reverse transcription reaction.• optimal choice for construction of cDNA libraries from eukaryotic mRNAs.• Full length cDNA cloning• 3’ rapid amplification of cDNA ends (3’ RACE)• Generation of labelled cDNA for screening of microarrays.Oligo (dT)20 Primer can not be used together with degraded RNA, prokaryotic RNA or miRNA (lack of poly(A) tail). Primer Sequence: 5´ – d (TTT TTT TTT TTT TTT TTT TT) –3´

Random Hexamers are short oligodeoxyribonucleotides of random sequence [d(N)6]. Primers are quality controlled (MS), purified (reversed phase HPLC), lyophilised and aliquoted and came with a guaranteed amount of at least 5OD (about 76.1nmol (136.4µg)). Hexanucleotide primers are a mixture of random 5'-hydroxyl hexanucleotides or hexamers, and can be used to quickly and efficiently prepare radioactive or non-radioactive probes using a DNA polymerase and a suitable DNA template or for cDNA synthesis from mRNA. The heterogeneous nature of the random primers ensures that all possible sequences will be represented in the probe mixture. As a result, these primers are well suited for generating as long as possible (complete) transcripts of the RNA, transcribing the 5 'ends, in particular for long RNA templates. As an alternative you can also use Oligo(dT)-Primer > for reverse transcription. Oligo(dT) primer bind to the polyA tail of RNA and therefore transcription will start always at the 3'-end of the RNA. The guaranteed delivery quantity of 5OD, corresponding to approx. 76.1nmol, or 136.4μg, can be dissolved in 760μL DEPC water to give a concentration of 100pmol/μL (100μM), or approx. 180μg/μL. If using between 50ng and 250ng of the hexamers per 20μL reverse transcription reaction, 760μL (100μM) will be sufficient for 460 to 2280 reverse transcriptase reactions.

The Genaxxon RNase Inhibitor is a recombinant (E.coli) murine protein which inhibits ribonucleases (RNases) A, B and C. It does not inhibit RNase 1, RNase H, RNase T1, S1 Nuclease. Native RNase-Inhibitor from murine placenta exerts its inhibitory effect by binding non-covalently to RNases in a 1:1 ratio with high affinity. There is no inhibition of polymerase activity when this RNase Inhibitor is used with AMV Reverse Transcriptase, M-MuLV Reverse Transcriptase, Taq DNA Polymerase or Phage RNA Polymerase (SP6, T7, or T3).The enzyme is active over a broad pH range between 5 and 9, with a maximum activity at pH7 – pH8. The inhibition of RNases is a reversible process. Reagents as urea or thiol reagents will lead to the dissociation of the Inhibitor-RNase-complex, resulting in re-natured and functional RNase and irreversibly deactivated RNase inhibitor.Applications: RT-PCRcDNA synthesisIn vitro transcription/translationEnzymatic RNA labelling reactionsIn vitro virus detection. Improvement of RNA translation in homologous systems.Preparations of RNase-free antibodies. Any application where eukaryotic RNase contamination is a potential problem.  Advantages:  The recombinant murine RNase inhibitor does not contain the cysteine pair that was identified to cause the high sensitivity against oxidation resulting in the inactivation of the human RNase inhibitor (1). As a result, the murine RNase inhibitor has significantly improved oxidation resistance compared to the human and porcine RNase inhibitors. In addition, murine RNase inhibitor is much more stable at low low DTT concentrations (less than 1 mM), what makes it ideal for reactions where high DTT concentrations interfere with the reaction (e.g. real-time RT-PCR). Product does not contain glycerol => lyo-ready product (ready for preparations of freeze-dried products).Recommendations: Since the RNase inhibitor protein is sensitive to denaturing agents and conditions, whereas RNases are not, care must be taken to avoid high concentrations of urea or other denaturing agents and temperatures above 50-55°C. The recommended concentration of the RNase inhibitor is 1 unit/µL. During set-up of a reaction, RNase inhibitor should be added before other components are added (enzymes, plasmids from mini preps, etc.).References1. Kim, B.M. et al. Protein Science. 8, 430-434.

The Genaxxon M-MuLV Reverse Transcriptase, encoded by Moloney Murine Leukemia Virus (M-MuLV RT) and expressed in E.coli is an RNA-dependent DNA polymerase lacking the RNase H activity. The enzyme synthesizes the cDNA first strand from a single-stranded RNA template to which a primer has been hybridized. M-MuLV RT will also extend primers hybridized to single-stranded DNA! The enzyme is recommended for synthesis of cDNA from 100 bp up to 10 kb length. Second strand cDNA synthesis can be achieved from some mRNA templates without an additional DNA polymerase. Genaxxon offers the HotScriptase master mixes (M3062 >; M3064 >) with which it is possible to synthesize double stranded DNA completely without an isothermal transcription step. Time saving: up to 50%! No need for Superscript!Money saving: high! With our high quality dNTPs as Set (M3015.4100 and M3015.0250) or Mix (M3016.1010) or our DNA Ladders and our favourable standard agarose (M3044) we can offer additional high quality products for your PCR.