Produits biochimiques et réactifs

<p>RNase H is a 18.9 kDa recombinant endoribonuclease purified from an Escherichia coli strain, which over-expresses cloned RNase H gene (rnh). The enzyme hydrolyses specifically the phosphodiester bonds of RNA hybridized to DNA and produces 5´ phosphate-terminated oligoribonucleotides and single-stranded DNA.<br> Rnase H does not degrade single and double-stranded DNA or unhybridized RNA. It is a key enzyme in the removal of mRNA after first-strand cDNA synthesis. Treating cDNA with RNase H prior to PCR can improve sensitivity as RNA bonded to the cDNA template may prevent binding of the amplification primers in a PCR reaction. RNase H treatment is often necessary when amplifying longer, full-length cDNA targets. In addition, RNase H is useful for the removal of poly(A) tails on mRNAs after hybridization with oligo(dT) and also for the site-specific enzymatic cleavage of RNA. </p>

T7 RNA polymerase is a monomeric bacteriophage encoded DNA directed RNA polymerase which catalyzes the formation of RNA in the 5"-3" direction.In the process of initiation of transcription T7 recognizes a specific promoter sequence, the T7 promoter.T7 consists of 883 amino a cids and has a molecular weight of 99 kDa.T7 RNA polymerase is isolated from E. coli BL21 carrying the plasmid pAR1219 which contains T7 gene I under the control of the inducible lac UV5 promoter.Concentration: 50,000units/ml (50U/µl)

The Water, nuclease-free, is deionized and 0.22 µm membrane-filtered. It is suitable for use in all molecular biology applications.

<p>This primer is a synthetic single-stranded 15-mer oligonucleotide with 5'- and 3'-hydroxyl ends. The primer is supplied as an aqueous solution and can be used for first strand cDNA synthesis synthesis in presence of reverse transcriptase. Oligo (dT) is one of the three primers that can be used during cDNA synthesis. It is used to prime all the mRNAs having a polyA 3' tail.</p>

Water is deionized, diethylpyrocarbonate (DEPC) treated and 0.22 µm membrane-filtered. It is recommended to be used in any application where RNA is involved

The set contains six ready-to-use 10X Taq Buffers with KCl for optimization of magnesium ion concentration in PCR applications with Taq DNA Polymerase. The MgCl2 concentration in each of the six buffers is different, ranging from 12.5mM to 30 mM.

10X Roseate PCR Buffer (with MgCl2) is recommended for all PCR applications with Taq DNA Polymerase (both recombinant and native)

TRIzol-C Reagent is our new and cost-effective ready-to-use formulation for isolation of pure total RNA from cells and tissues. The reagent, a mono-phasic solution of phenol and guanidine isothiocyanate, is an improvement giving very high yields of pure RNA useful for downstream processes . During sample homogenization or lysis, TRIzol-C Reagent maintains the integrity of the RNA, while disrupting cells and dissolving cell components. Addition of chloroform followed by centrifugation, separates the solution into an aqueous phase and an organic phase. RNA remains exclusively in the aqueous phase. After transfer of the aqueous phase, the RNA is recovered by precipitation with isopropyl alcohol. After removal of the aqueous phase, the DNA and proteins in the sample can be recovered by sequential precipitation. Precipitation with ethanol yields DNA from the interphase, and an additional precipitation with isopropyl alcohol yields proteins from the organic phase. Copurification of the DNA may be useful for normalizing RNA yields from sample to sample.

10X PCR Buffer with KCl (MgCl2+ free) is recommended for all PCR applications with Taq DNA Polymerases (both recombinant and native).

PCR Enhancer System is used in conjunction with DNA polymerases to optimize PCR of problematic and/or GC-rich templates. The system offers higher primer specificity, broader magnesium concentration, broader annealing temperature,and improved themostability DNA polymerases.

RT-PCR Kit is optimized to synthesize first-strand cDNA from purified poly (A)+ or total RNA. The RT-PCR kit for RT-PCR delivers increased cDNA yields, high sensitivity, and full-length transcripts in a convenient format. All the components needed for successful first-strand cDNA synthesis is included, saving time and ensuring success with every experiment. The kit Reverse Transcriptase is a version of M-MLV RT that has been engineered to reduce RNase H activity and provide increased thermal stability. The enzyme is used to synthesize cDNA at a temperature range of 42-55°C, providing increased specificity, higher yields of cDNA, and more full-length product than other reverse transcriptases.

SDS-PAGE is a common technique used to study the electrophoresis of proteins. Preparation of polyacrylamide gels is cumbersome and gel strength is based on accurate additions of reagents. To simplify this process and eliminate pipetting errors, ready-to-use Stacking gel mix as a 4% solution mix is offered.