recombinant RNase Inhibitor - RNasin

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´

Ribonuclease A is an endo-ribonuclease specifically cleaving single stranded RNA at the 3' side of pyrimidine bases (cytosine and uracil). RNAse A ist used to prepare RNA-free DNA, to digest non-hybridised regions of RNA-DNA hybrids and as a molecular weight marker. The pH optimum for RNAse A is between 7.0-7.5. RNAse A is inhibited by Diethylpyrocarbonate (DEPC) >, guanidinium salt (4M GuaSCN), beta-Mercaptoethanol, heavy metals, vanadyl-ribonucleoside complexes, RNAse-inhibitor from human placenta and by competitive DNA. RNAse A cleaves single- and double-stranded RNA and RNA in RNA:DNA hybrids at low salt conditions (100 mM NaCl). At high salt conditions (>300 mM NaCl), RNAse A cleaves single-stranded RNA only. The enzyme can only be removed by Proteinase K digest and subsequent phenol extraction. Elimination of DNAse activity: Product S5231 (RNAse A (not certified DNAse free)) may contain DNAse activity. As RNAse A is heat stable it is recommended to heat inactivate DNAse activity before use. Procedure: 10mg/mL RNAse A in 0.01 sodium acetate (pH5.2) have to be heated to 100°C for 15 minutes in a water bath. After 15 minutes, the RNAse shall remain in the water bath while the water cools down to room temperature. pH can be adjusted by addition of 0.1 time the volume of 1M Tris-HCl (pH7.4). After aliquotation of the now DNAse free RNAse A solution, each aliquote should be stored a -20°C. NOTE: RNAse A precipitates if concentrated solutions are heated to 100°C at neutral pH! Stock solution: Stock solutions are prepared at concentrations from 1 - 10mg/mL in 10mM Tris/HCl, pH7.5; 15mM NaCl or in 10mM Tris/HCl, pH7.5; 1mM EDTA, pH8.0 (TE buffer).The recommended working concentration is 10µg/mL (removal of RNA from plasmid preparations; 1hr, RT) or 100ng/mL (preparation of 'blunt ends' of double-stranded cDNA). Stability:RNase A aggregates during lyophilizing and storage. It has a high affinity to glas surfaces, which has to be taken into consideration. At neutral pH (e. g. in PBS pH 7.4) and high concentrations (>10mg/mL) the enzyme will precipitate. At +2°C to +8°C (lyophilized) it is stable for several years (dry storage), in solution (-20°C) several years or (+2°C to +8°C) several weeks.

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 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.

Ribonuclease A- DNase-free is a DNase and Protease-free enzyme that can be used directly for RNA digests, e.g. in DNA purification procedures. Elimination of potential DNAse activity is not necessary. Potential DNAse activities have been removed. DescriptionRibonuclease A is an endo-ribonuclease specifically cleaving single stranded RNA at the 3' side of pyrimidine bases (cytosine and uracil). RNAse A ist used to prepare RNA-free DNA, to digest non-hybridised regions of RNA-DNA hybrids and as a molecular weight marker. The pH optimum for RNAse A is between 7.0-7.5. RNAse A is inhibited by Diethylpyrocarbonate (DEPC) >, guanidinium salt (4M GuaSCN), beta-Mercaptoethanol, heavy metals, vanadyl-ribonucleoside complexes, RNAse-inhibitor from human placenta and by competitive DNA. RNAse A cleaves single and douple stranded RNA and RNA in RNA:DNA hybrids at low salt conditions (100mM NaCl). At high salt conditions (>300 mM NaCl), RNAse A cleaves single-stranded RNA only. The enzyme can only be removed by Proteinase K digest and subsequent phenol extraction. Stock solution: Stock solutions are prepared at concentrations from 1 - 10mg/mL in 10mM Tris/HCl, pH7.5; 15mM NaCl or in 10mM Tris/HCl, pH7.5; 1mM EDTA, pH8.0 (TE buffer).The recommended working concentration is 10µg/mL (removal of RNA from plasmid preparations; 1hr, RT) or 100ng/mL (preparation of 'blunt ends' of double-stranded cDNA). Stability:RNase A aggregates during lyophilizing and storage. It has a high affinity to glas surfaces, which has to be taken into consideration. At neutral pH (e. g. in PBS pH 7.4) and high concentrations (>10mg/mL) the enzyme will precipitate. At +2°C to +8°C (lyophilized) it is stable for several years (dry storage), in solution (-20°C) several years or (+2°C to +8°C) several weeks.