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Custom and library siRNA for efficient gene silencing

High-quality short interfering RNA (siRNA) from QIAGEN allows efficient gene silencing in eukaryotic cell culture by targeting complementary endogenous mRNA. This effect, termed RNA interference (RNAi), enables targeted gene silencing in functional genomics studies. Now, QIAGEN offers a custom siRNA oligonucleotide synthesis service that includes a range of modifications and also library siRNAs directed against common target genes.

QIAGEN siRNA Oligo Synthesis Service offers:
  • High-purity siRNA for efficient gene silencing
  • Custom and library siRNA ready to use in cell transfection
  • Expert advice on siRNA design
  • Labeled siRNA
Multiplex PCR that simply works - the new QIAGEN Multiplex PCR Kit

The new QIAGEN Multiplex PCR Kit is the first kit specifically developed for multiplex PCR. The simple multiplex master-mix solution eliminates the need for lengthy optimization procedures, such as adjusting the amounts of Mg2+ and enzyme or even, as frequently required, adjusting primer concentrations. Now standard multiplex PCR applications are fast and easy to perform.

Advantages of the new QIAGEN Multiplex PCR Kit:
  • No optimization required - easy assay development with simple master mix
  • High specificity and sensitivity - stringent hot start with HotStarTaq DNA Polymerase and increased sensitivity with the unique new multiplex PCR buffer
  • Versatile for many applications - including typing of tran sgenic animals and plants, detection of bacteria and viruses, and microsatellite analyses
  • Easy to use and cost-effective - simple reaction setup for fast and reproducible result
Principle of siRNA-mediated RNA interference

The application of RNAi technology to mammalian cells has revolutionized the field of functional genomics. Use of siRNA allows targeted inhibition of gene expression using double-stranded RNA (dsRNA) that carries the same sequence as mRNA transcribed from the target gene. In mammalian cell systems, these double-stranded siRNA oligos can be used to induce gene silencing. Targeting a specific gene by RNAi requires a double-stranded siRNA containing the sense and antisense sequence of mRNA transcribed from the target gene. QIAGEN uses patented TOM-amidites to synthesize high-quality, highpurity, 21-nucleotide sense and antisense RNA oligonucleotides. In mammalian cells, siRNA must be delivered to the cytoplasm using an RNA transfection reagent. TransMessenger Transfection Reagent from QIAGEN efficiently delivers siRNA to cells for successful gene knock down (Figures 1 and 2). Once inside the cell, current models suggest that siRNA interacts with endogenous cellular proteins to form a proteinRNA complex known as the RNA induced silencing complex (RISC). Due to the sequence homology between the siRNA and the target mRNA, the RISC specifically targets the mRNA for enzymatic degradation (see flowchart).

Efficient Gene Silencing Using QIAGEN siRNA and TransMessenger Reagent siRNA-Mediated RNAi Figure 1 Western blot of extracts from transfected HeLa-S3 cells probed with lamin A/C monoclonal primary
antibodies. Cells were transfected using the indicated reagents. siRNA: short interfering RNA duplex targeting
lamin A/C at position 608630 relative to the start codon; TM: TransMessenger Transfection Reagent; ODN:
Scrambled 24mer oligodeoxynucleotide (control). Efficient, Targeted Gene Silencing Figure 2 Relative expression levels of lamin A/C after siRNA transfection. The value for the mock transfection was set as 1. Data were normalized to the expression level of GAPDH (internal control), which was quantified
by real-time RT-PCR using OmniscriptReverse Transcriptase and the QuantiTect SYBR Green PCR Kit. Expression levels were calculated using a standard curve generated using different amounts of HeLa S3 cDNA. Data obtained under the indicated transfection conditions are shown as relative, normalized expression levels. Total RNA was prepared using the RNeasy Mini Kit and the QIAShredder homogenizer.
High-quality siRNA for efficient gene silencing

QIAGEN offers two grades of custom siRNA; ultrapure siRNA purified by ion-exchange HPLC (>97% pure), and crude siRNA (8085% pure). In addition, a number of library siRNA duplexes (>97% pure) are available for silencing common target genes. All siRNA products are delivered annealed,* desalted, deprotected, and ready to use. A detailed certificate of a nalysis, including HPLC analysis, is included with each order. All QIAGEN siRNA is supplied with a sterile resuspension buffer and is suitable for use in transfection procedures.

* Non-annealed siRNA is available on request.
HPLC trace of the siRNA duplex is provided. If non-annealed siRNA is requested, two individual chromatograms of the single strands are provided.



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