GSC Human Pre-designed siRNA Set A

Cat. No.: G1479927

Storage

Store at -20°C

Shipped In

Ice chest + Ice pads

Panoramica

GSC Human Pre-designed siRNA Set A contains three designed siRNAs for GSC gene (Human), as well as a negative control, a positive control, and a FAM-labeled negative control. Components GSC siRNA-1: 5 nmol (HPLC) GSC siRNA-2: 5 nmol (HPLC) GSC siRNA-3: 5 nmol (HPLC) siRNA Negative Control: 5 nmol (HPLC) FAM-labeled siRNA Negative Control: 5 nmol (HPLC) GAPDH siRNA Positive Control:5 nmol (HPLC)

Specifications

Condizioni di conservazione di stoccaggio

Store at -20°C

Spedito in

Ice chest + Ice pads

Questo prodotto richiede spedizione a catena fredda. I servizi di terra e altri servizi economici non sono disponibili.

Product Line

Prodotti biologici

Application Protocols

Tested applications, recommended working concentrations, and validated cell lines for SKU G1479927:

• Not specified for this item; refer to CoA/Spec Sheet and SDS.

General recommendation: Follow your institution’s standard RNAi transfection SOPs and the instructions of the chosen transfection reagent. Perform small-scale optimization for dose, reagent ratio, and timing prior to large experiments.

Biological Roles

General/literature context (not item-specific):

• siRNA function: exogenous siRNAs exploit the endogenous RNA interference pathway to downregulate gene expression post-transcriptionally. The guide strand directs Argonaute (AGO2) to complementary mRNA, resulting in catalytic cleavage and subsequent exonucleolytic decay.
• Cellular processes impacted: gene-specific knockdown enables dissection of signaling pathways, cell cycle control, apoptosis, differentiation, and metabolic regulation.
• Innate immune considerations: certain sequences or unmethylated motifs can trigger innate sensors (TLR3, RIG-I, MDA5). 2′-O-methyl or other modifications in seed/uridine-rich regions can mitigate this response in some systems.
• Off-targets: microRNA-like seed-mediated effects can downregulate unintended transcripts. Use multiple independent siRNAs and rescue experiments to confirm on-target biology.
• Duration of effect: typically transient (days); persistence depends on cell division rate, nuclease activity, and sequence/chemical stabilization.

Item-specific biological targets, sequences, or modifications: Not specified for this item; refer to CoA/Spec Sheet.

Buffer Applications

While this is not a buffer chemical, appropriate buffer use is essential for siRNA stability and delivery.

General guidance (literature; not item-specific):

• Stock preparation: dissolve lyophilized siRNA in RNase-free water or TE buffer to 10–100 μM. TE (10 mM Tris-HCl, 1 mM EDTA, pH 7.5–8.0) enhances stability for long-term storage; dilute EDTA below 0.1 mM prior to complex formation with cationic lipids/polymers.
• Working buffers: follow transfection reagent recommendations (often reduced-serum media like Opti-MEM for complex assembly). Final transfection mixes are typically returned to complete growth media after complexation.
• pH/salt: maintain physiological osmolarity and pH ~7.2–7.4 for cell culture. Avoid Mg2+/Ca2+ during prolonged storage without chelation due to nuclease activation.
• Sterility: use sterile, nuclease-free buffers; filter (0.22 μm) if needed, taking care to minimize nucleic acid adsorption to filters.

Item-specific buffer composition or provided solvent: Not specified for this item; refer to CoA/Spec Sheet.

Green Alternatives

This product is an oligonucleotide reagent; green chemistry considerations focus on minimizing solvent use, hazardous reagents in delivery, and waste in high-throughput workflows.

Greener practice suggestions (general; not item-specific):

• Prefer aqueous preparation: reconstitute and dilute exclusively in RNase-free water or TE buffer, avoiding organic co-solvents.
• Delivery systems: choose lower-toxicity, serum-compatible transfection reagents that permit reduced reagent volumes, minimizing waste and cell stress.
• Miniaturization: adopt microplate miniaturization (384/1536-well) with acoustic or low-dead-volume dispensing to reduce reagent consumption and plastic waste.
• Workflow design: multiplex readouts to decrease repeat transfections; implement robust positive/negative controls to avoid reruns.
• Synthesis impacts: where options exist, select oligos manufactured with greener phosphoramidite processes and solvent recovery (supplier dependent; consult sustainability statements).

Illustrative comparison (general):

• Conventional: lipid transfection in serum-free media with multiple washes; high volumes and plastic use.
• Greener: serum-compatible formulations allowing direct add-in, single-step protocols, and reduced media exchanges.

Note: No item-specific environmental certifications or solvent data are provided for SKU G1479927; refer to CoA/Spec Sheet or SDS if available.

Pharmaceutical Uses

No clinical or therapeutic claims are made for this product. For research use only.

General formulation context for siRNA in research (not item-specific):

• Discovery workflows: siRNA sets are widely used for in vitro target validation, lead prioritization, and mechanism studies prior to any drug development.
• Excipients in research formulations: cationic lipids, polymers (e.g., PEI), or lipid nanoparticles are commonly used carriers for cell-based delivery; selection impacts potency and cytotoxicity.
• Quality attributes considered in regulated settings (context only): identity, purity, endotoxin, residual solvents, and nuclease stability; however, no pharmacopeial monographs apply directly to sequence-defined siRNA mixtures used as research tools.

Item-specific pharmacopeial status, GMP grade, or excipient role: Not specified for this item; refer to CoA/Spec Sheet.

Physical Properties

Item-specific physicochemical specifications (BP/MP/density/UV cutoff/logP, etc.): Not specified for this item; refer to CoA/Spec Sheet.

General/literature properties of siRNA (for context; not item-specific):

• Physical state: lyophilized solid or aqueous solution, depending on packaging; hygroscopic and moisture-sensitive to RNases.
• Solubility: readily soluble in nuclease-free water, TE buffer (10 mM Tris-HCl, 1 mM EDTA, pH 7.5–8.0), or PBS for cellular delivery. Avoid divalent cations without chelators to limit nuclease activity.
• pKa/logP: Not applicable as discrete values; polyanionic macromolecules with multiple phosphate groups; effectively highly hydrophilic.
• UV absorbance (literature): strong at 260 nm; typical conversion A260 = 1.0 ≈ 40 μg/mL for single-stranded RNA, duplexes similar but sequence-dependent. Use supplier-provided OD for exact quant.
• Thermal behavior: duplex melting temperature (Tm) depends on length, GC content, salt concentration, and chemical modifications; typical 21-mer siRNA Tm often >65 °C in physiological salt but varies widely.
• Stability: sensitive to hydrolysis and RNases; improved by lyophilization, low-temperature storage, and optional backbone/sugar modifications.

Notes:

• Do not treat the above values as specifications for SKU G1479927. For any numeric guarantees, consult the product CoA/Spec Sheet.

Quality and Grades

Item-specific grade/purity details: Not specified for this item; refer to CoA/Spec Sheet.

Context for siRNA reagent quality (general, not item-specific):

• Common purification levels:
  • Desalted: removal of salts/small-molecule impurities; adequate for many cell-based assays but may contain n-1 species.
  • HPLC-purified: higher purity; reduces truncated sequences; preferred for demanding phenotypes or in vivo-like models.
  • PAGE-purified: very high length homogeneity; often used for precise studies.
• Optional quality attributes: strand identity confirmation by LC–MS or MALDI, OD260 and nmol content reporting, endotoxin testing (for sensitive primary cells), residual solvent/salt analysis.
• Stabilizers/modifications: 2′-OMe/2′-F bases, phosphorothioate linkages, or terminal capping can enhance nuclease resistance and potency. Presence/absence of such modifications for this item is Not specified; refer to CoA/Spec Sheet.
• What “pre-designed set” typically implies (literature): multiple independent siRNA duplexes per target gene to ensure robust knockdown and mitigate off-target effects; sequences are bioinformatically filtered for low off-target potential and appropriate thermodynamic asymmetry.

Recommendation: For publication-critical work, request and retain the CoA including purification method, OD/nmol content, and endotoxin result where relevant.

Reaction and Applications

As a functional genomics tool, GSC Human Pre-designed siRNA Set A is intended for RNA interference (RNAi)-mediated transient gene silencing in human cells.

General/literature applications (not item-specific):

• Gene knockdown: sequence-specific depletion of mRNA to interrogate gene function, epistasis, and pathway mapping.
• Assay integration: phenotypic screens (imaging, viability, reporter assays), target validation prior to CRISPR knockout, synthetic lethality studies, and mechanism-of-action deconvolution.
• Co-delivery: combine with plasmids, reporters, or chemical probes to assess pathway dependencies.
• Controls: use non-targeting siRNA, positive control siRNA (e.g., PLK1, KIF11) for transfection potency, and siRNA-resistant rescue constructs for on-target verification.

Mechanistic notes:

• siRNA duplex is loaded into RISC; the passenger strand is discarded, and the guide strand directs Argonaute-mediated endonucleolytic cleavage of complementary mRNA.
• Efficacy determinants: sequence design, seed-region off-target minimization, GC content (~30–52% typical), thermodynamic asymmetry, and chemical stabilization for nuclease resistance.

Practical tips:

• Use RNase-free technique; thaw on ice, briefly spin, and mix gently.
• Optimize transfection conditions (cell density, reagent type, charge ratio, and serum presence) for each cell line.
• Time course: maximal knockdown typically 24–72 h post-transfection; proteins with long half-lives may require 96 h or repeat dosing.

Item-specific application claims: Not specified; refer to CoA/Spec Sheet.

Reaction Conditions

Classical reaction conditions do not apply; however, successful siRNA-mediated knockdown depends on optimized transfection conditions. The following are general, literature-based guidelines (not item-specific specifications):

• Cell density: seed cells to reach 40–70% confluence at transfection time (adherent lines); optimize per cell type.
• siRNA dose: typical final 1–50 nM per duplex; use the lowest effective concentration to reduce off-targets. For pooled sets, maintain constant total siRNA by reducing individual duplex amounts proportionally.
• Complex formation: mix siRNA with transfection reagent in serum-free or reduced-serum medium; incubate 5–30 min to allow complexation before adding to cells. Follow vendor ratios for each reagent.
• Incubation: return to complete medium after 4–6 h if the reagent requires; many modern reagents allow no-change protocols.
• Timing of readouts: assess mRNA knockdown at 24–48 h (qPCR) and protein depletion at 48–96 h (WB/IF/assay), depending on protein half-life.
• Repeat dosing: for long-lived proteins or rapidly dividing cells, a second dose at 48–72 h may enhance silencing.
• Controls: include non-targeting siRNA, positive control siRNA, and transfection-reagent-only controls; verify knockdown with orthogonal assays.

Item-specific recommended conditions or verified cell lines: Not specified for this item; refer to CoA/Spec Sheet.

Safety and Handling

• GHS/CLP classification, signal word, pictograms, H-statements: Not specified for this item; refer to SDS for authoritative safety information.
• Research use: For research use only (non-clinical, non-diagnostic).

General laboratory safety for synthetic siRNA reagents (literature/good practice):

• Hazards: siRNAs are non-hazardous under typical lab handling; the principal risks are contamination with RNases and potential biological activity in cells. Handle using aseptic, RNase-free technique.
• PPE: lab coat, gloves (nitrile), safety glasses. Change gloves frequently to avoid RNase transfer; consider RNase decontaminants for surfaces and tools.
• Handling: work in clean areas; use RNase-free consumables and solutions. Keep on ice during setup; minimize freeze–thaw cycles by aliquoting.
• Incompatibilities: avoid exposure to RNases (human skin, non-sterile plastics), high pH (>9) and prolonged elevated temperature. Avoid divalent cations without chelation (can enhance nuclease activity).
• First aid (general): in case of skin/eye contact, rinse with water. If inhaled (powder), move to fresh air. If ingested, rinse mouth with water. Seek medical attention as needed. Refer to SDS for details.
• Waste: dispose per institutional biosafety guidelines. Though non-infectious, treat as bioactive nucleic acid; decontaminate surfaces with RNase-inactivating agents.
• Transport: shipped in insulated packaging with ice packs; avoid temperature excursions.

Solvent Selection

This product is an aqueous biopolymer reagent rather than an organic solvent; classical solvent selection is not applicable. Instead, focus on reconstitution and handling buffers compatible with RNA and transfection.

General buffer guidance for siRNA (literature/good practice):

• Primary diluents: RNase-free water or TE buffer (10 mM Tris-HCl, 1 mM EDTA, pH 7.5–8.0). TE improves long-term stability via chelation of divalent cations.
• Working solutions: RNase-free water or buffer provided by the transfection reagent manufacturer (e.g., Opti-MEM for complex formation when specified). Avoid serum during complexation if required by the reagent.
• Ionic strength/pH: maintain near-neutral pH (7.0–8.0). High ionic strength can influence duplex stability and complex formation; follow the transfection reagent’s instructions.
• Organic solvents: avoid DMSO/DMF for stock solutions; siRNA is fully water-soluble. If delivered in DMSO-dependent screening workflows, keep DMSO ≤1–2% v/v in final cell media to limit cytotoxicity (when absolutely necessary).

Quick comparison (general):

• RNase-free water: maximal compatibility; ideal for short-term use and transfection complexation.
• TE buffer: improved storage stability; EDTA may chelate cations used by some delivery systems—dilute sufficiently before mixing with cationic lipids/polymers.

Item-specific buffer recommendations: Not specified for this item; refer to CoA/Spec Sheet.

Storage and Reconstitution

• Item-specific storage: Store at -20°C.
• Shipping: Shipped in ice chest with ice pads to maintain low temperature.
• Research use note: For research use only.

General/literature guidance for siRNA handling (not item-specific):

• Upon receipt: If lyophilized, briefly centrifuge to collect material. Reconstitute in RNase-free water or TE buffer to 10–100 μM. If in solution, verify concentration and buffer.
• Aliquoting: Prepare single-use aliquots to avoid freeze–thaw; label with sequence ID/lot/date. Avoid more than 3 freeze–thaw cycles.
• Storage conditions: Short term (≤1 week) at 4°C; medium term at -20°C; long term at -80°C for maximum stability. Protect from repeated warming.
• Light/moisture: Store in low-humidity, protected from light if fluorescent labels are present. Use nuclease-free, low-binding tubes.
• Thawing: Thaw on ice, mix gently (do not vortex vigorously), quick spin before opening to collect condensate.
• Working solutions: Keep on ice during setup; discard leftovers rather than refreezing when possible.

Item-specific reconstitution solvent, concentration, or stabilizers: Not specified for this item; refer to CoA/Spec Sheet.

Structure and Identity

This product is a set of pre-designed small interfering RNAs (siRNAs) for human gene silencing applications.

• Item-specific identifiers:

  • SKU: G1479927
  • Product name: GSC Human Pre-designed siRNA Set A
  • CAS, CID, InChIKey, SMILES: Not specified for this item; refer to CoA/Spec Sheet.
  • Molecular formula, Molecular weight: Not specified for this item; refer to CoA/Spec Sheet.

• General/literature description of siRNA (not item-specific):

  • siRNAs are short double-stranded RNA duplexes typically ~21–23 nt with 2-nt 3′ overhangs on each strand.
  • Each duplex comprises an antisense (guide) strand complementary to the target mRNA and a sense (passenger) strand.
  • Common chemical architecture: 5′-phosphate/5′-OH termini; 3′-UU or dTdT overhangs; optional stabilizing modifications (e.g., phosphorothioates, 2′-O-methyl, 2′-F) depending on design.
  • Structural features: A-form RNA helix; Watson–Crick base pairing; possible sequence-dependent thermodynamic asymmetry to favor RISC loading of the guide strand.

• 2D structural description (general):

  • Two antiparallel ribonucleotide strands (ribose-phosphate backbone) with canonical A/U and G/C base pairs; 3′ overhangs protrude as short single-stranded tails. No stereogenic centers beyond ribose configuration (D-ribose).

Synthetic Utility

This product is a functional genomics reagent (siRNA), not a synthetic building block or catalyst. It is not typically used as a reagent for chemical synthesis.

For chemists integrating biology with synthesis (general guidance):

• Use siRNA knockdown to validate biosynthetic enzyme targets before designing chemical inhibitors.
• Couple phenotypic outcomes with chemoproteomics to link small-molecule engagement to gene function.

Item-specific synthetic reactivity or transformations: Not applicable.

Target Specificity

Item-specific details such as target gene(s), siRNA sequences, pool composition, off-target filtering criteria, and cross-reactivity:

• Not specified for this item; refer to CoA/Spec Sheet.

General note (not item-specific): Pre-designed siRNA sets typically include multiple independent duplexes per target to ensure on-target verification and enable pooled or individual testing to mitigate seed-based off-target effects.

Domande frequenti

How should this product be stored?

Store at ?20 °C. Freezer storage is required to maintain the specified shelf life.

How is this product shipped?

This product ships in an insulated container with ice pads. Unpack on arrival and transfer it to the storage condition stated above.

What documentation is provided?

Available product documentation, including Certificates of Analysis (COA), Safety Data Sheets (SDS), and specification sheets, is shown in the product document area. Document availability and access follow the current site policy.