Human PPIH qPCR Primer Pair

Cat. No.: H746972

Storage

Store at -20°C,Avoid repeated freezing and thawing

Shipped In

Ice chest + Ice pads

Spezifikationen

Stabilität und Lagerung

Store at -20 ℃. Suggest appropriate packaging after reconstitution to avoid repeated freeze-thaw cycles.

Storage

Store at -20°C,Avoid repeated freezing and thawing

Verschickt in

Ice chest + Ice pads

Dieses Produkt erfordert Kühlkettenversand. Grundversand und andere Economy-Optionen sind nicht verfügbar.

Product Line

Biologische Produkte

Application Protocols

Item-specific validated protocols

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

General qPCR workflow (literature guidance; adapt to your master mix/instrument)

1. Primer reconstitution and dilution

• Spin tubes briefly. Reconstitute each primer in nuclease-free water or low-TE to 100 µM. Vortex gently and incubate 10 min to ensure dissolution. Prepare 10 µM working aliquots and store at −20 °C.

2. Reaction setup (example 20 µL)

• 10 µL 2× qPCR master mix (SYBR Green or probe-based)
• 0.4–1.0 µL 10 µM Forward primer (final 200–500 nM)
• 0.4–1.0 µL 10 µM Reverse primer (final 200–500 nM)
• x µL template cDNA (amount optimized; e.g., equivalent to 1–10 ng RNA)
• Nuclease-free water to 20 µL
• Include NTC and no-RT controls.

3. Cycling (example)

• 95 °C 2–3 min; 40 cycles of 95 °C 10–15 s, 58–62 °C 20–30 s. Run melt curve if using SYBR chemistry.

4. Data analysis

• Confirm single melt peak and expected Ct range. Generate standard curve to calculate efficiency. Normalize PPIH Ct to chosen reference genes using ΔΔCt where appropriate.

Notes

• Optimize annealing temperature via gradient if non-specific amplification or primer–dimer is observed. Exact Tm/amplicon details must be taken from this product’s CoA once available.

Biological Roles

Item-specific context

• Target: Human PPIH (Peptidylprolyl isomerase H), also known as Cyclophilin H. This primer pair is intended to amplify PPIH transcripts for qPCR-based analysis.

General biology of PPIH (literature)

• Function: PPIH is a cyclophilin-family peptidyl-prolyl cis-trans isomerase implicated in protein folding and RNA splicing machinery. It associates with components of the spliceosome (e.g., PRPFs) and may facilitate conformational rearrangements during spliceosome assembly.
• Localization: Predominantly nuclear with spliceosomal enrichment reported; may shuttle depending on cell state.
• Pathways: Involved in pre-mRNA processing/spliceosome pathway; part of broader protein-folding/isomerase functional network.
• Expression: Generally ubiquitous but with cell-type and condition-dependent regulation; serves as a candidate normalization gene in some contexts, though suitability must be experimentally confirmed.
• Orthology: Cyclophilins are conserved across eukaryotes; orthologs exist in many species with varying sequence identity.

Implications for qPCR assay design (general)

• Transcript complexity: Alternative splicing may produce multiple isoforms. Primer placement (exon–exon junctions, constitutive exons) will govern isoform coverage; sequence details for this product are not specified—consult CoA for exact amplicon mapping.
• Genomic DNA risk: Intronic regions around PPIH vary; DNase treatment and intron-spanning primers are recommended to minimize gDNA amplification.

Buffer Applications

Item-specific status

• No specific buffer formulation is provided for this item. Storage at −20 °C and avoidance of repeated freeze–thaw are indicated.

General guidance (literature)

• Reconstitution buffers: Nuclease-free water for immediate use; low-TE (10 mM Tris-HCl, 0.1–1 mM EDTA, pH 7.5–8.0) for improved long-term stability by chelating divalent cations that promote nuclease activity.
• Working solutions: Dilute primer stocks into nuclease-free water to prepare 10 µM working aliquots; add to qPCR mastermix, which supplies optimal Mg2+ and ionic strength.
• pH and ionic considerations: Maintain near-neutral pH. Avoid high-salt storage buffers that can precipitate upon freezing and complicate accurate pipetting at small volumes.
• Additives: Do not include PCR enhancers (e.g., DMSO, betaine) in the primer stock; introduce them only in the final reaction as needed.

Applications outside of qPCR

• While primarily for qPCR, the same primers may be used for endpoint PCR and melt-curve verification when validated; buffer conditions then follow standard PCR practice. Any such use should be validated empirically as sequences/amplicon parameters are not specified for this item.

Green Alternatives

Applicability

• For a qPCR primer pair, “greener” considerations focus on assay design, waste minimization, and energy use rather than solvent replacement.

Greener practices (literature/best practice)

• Reduce plastic waste: Use low-dead-volume tips, multi-well formats, and recycle outer packaging. Consolidate mastermix preparation to minimize consumables.
• Energy efficiency: Store working aliquots at −20 °C and bulk at −20 °C (per product guidance). Avoid unnecessary −80 °C storage. Use room-temperature stable qPCR master mixes when compatible to reduce cold-chain burden.
• Right-first-time design: Validate primer specificity in silico to reduce reruns and reagent waste.
• Miniaturization: Downscale reaction volumes (e.g., 10 µL or 5 µL) when instrument allows, cutting reagent and plastic consumption proportionally without sacrificing data quality.
• Non-hazardous reagents: Prefer intercalating dyes or probe chemistries with lower hazard profiles and avoid ethidium bromide gels for post-run checks; use safer nucleic acid stains.

Trade-offs

• More rigorous purification (e.g., HPLC) can improve performance and reduce repeats but has a higher synthesis footprint; balance with assay criticality.
• Single-use plastics ensure nuclease control but increase waste; consider certified nuclease-free reusable racks and responsible disposal streams.

Pharmaceutical Uses

Item-specific statement

• This product is designated For research use only. No pharmaceutical, diagnostic, or clinical use is claimed or supported.

General information (context only; not product claims)

• DNA oligonucleotides can serve as critical raw materials in regulated environments when appropriately qualified. However, qPCR primers for research are not formulated as excipients or active ingredients.
• If primers are used in GMP-adjacent workflows (e.g., in-process analytics), formal supplier qualification, change control, and lot-specific CoAs are typically required by quality systems.

Best practices for regulated labs

• Establish incoming inspection criteria: sequence verification, purity assessment, endotoxin/nuclease specifications if applicable to the process (not specified for this item).
• Maintain segregation of research-grade materials from GMP suites unless fully qualified and approved by QA.

Conclusion

• This item is intended strictly for laboratory research applications such as gene expression studies and method development, not for therapeutic manufacturing or patient-facing diagnostics.

Physical Properties

Item-specific properties (from Product Data)

• Appearance: Not specified for this item; refer to CoA/Spec Sheet.
• Molecular weight: Not specified for this item; refer to CoA/Spec Sheet (depends on primer sequences and length).

General/literature properties for DNA oligos (for context; not product specifications)

• Physical state: Typically supplied lyophilized (white solid) or in aqueous solution.
• Solubility: Freely soluble in nuclease-free water and TE buffer (10 mM Tris, 0.1–1 mM EDTA, pH ~7.5–8.0).
• Extinction coefficient (ε260): Sequence-dependent; used to quantify concentration by A260 (literature method).
• Melting temperature (Tm): Sequence- and salt-dependent. Short qPCR primers (18–25 nt) are usually designed for Tm ~58–65 °C (literature), enabling annealing ~58–62 °C with typical polymerases.
• pH stability: Stable near neutral pH; depurination accelerates at low pH and elevated temperature (literature).
• Divalent cations: Mg2+ stabilizes duplex formation; EDTA chelates divalents, protecting from nuclease activity (literature).

Practical notes

• Reconstitution concentration: Common working stocks 10–100 µM; preparation depends on intended qPCR setup (literature guidance).
• Quantification: UV A260 with sequence-specific ε or generic estimate (1 A260 unit ≈ 33 µg/mL ssDNA; literature).

Quality and Grades

Item-specific quality (from Product Data)

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

General guidance on primer quality (literature/industry practice)

• Synthesis quality: Oligos are commonly available as desalting-grade, cartridge/OPC-purified, HPLC-purified, or PAGE-purified. qPCR applications often perform well with standard desalting or cartridge-purified primers; high-GC or sensitive assays may benefit from HPLC/PAGE.
• Analytical characterization: Providers typically report sequence, yield, calculated molecular weight, and UV purity profile (A260/A280). Mass spectrometry (MALDI/ESI) can verify mass; capillary electrophoresis or HPLC can assess length distribution.
• UV background and contaminants: Residual small-molecule impurities or truncated sequences can impact amplification efficiency and specificity; purification level mitigates these.
• Batch traceability: For regulated labs, retain CoA, lot number, and synthesis report. Re-validate primer efficiency (standard curve) upon lot changes.
• Stabilizers/additives: Not specified for this item. Some suppliers add low-TE for liquid format or trehalose for stability; confirm on CoA if applicable.

What this means for this item

• Without a specified grade, users should verify performance in-house by determining amplification efficiency (ideal 90–110%), linear dynamic range, and specificity (single melt-curve peak) before critical experiments.

Reaction and Applications

Item-specific scope (from Product Data)

• Application: qPCR (quantitative PCR) primer pair targeting human PPIH. Additional validated applications are not specified for this item.

General applications and best practices (literature)

• Gene expression quantification: Use with cDNA templates to measure PPIH transcript abundance by SYBR Green or probe-based qPCR chemistries (primers applicable to both). Verify single amplicon by melt-curve and agarose gel if needed.
• Reference gene testing: PPIH is sometimes evaluated as a candidate reference gene; suitability is tissue/condition-dependent and must be empirically validated.
• Standard curves and efficiency: Generate a 5–7 point, 10-fold serial dilution curve to assess efficiency (ideal 90–110%) and linearity (R2 ≥ 0.99).
• Specificity optimization: Check for primer–dimer and off-targets using in silico tools (BLAST) against the human transcriptome/genome. Amplicon size of 70–200 bp is typical for qPCR (actual size not specified for this item).
• cDNA considerations: Use DNase-treated RNA and intron-spanning primers when possible to minimize genomic DNA amplification (design details not specified for this item).
• Controls: No-RT, NTC, and positive control sample recommended.

Practical tips

• Primer concentration: Common starting point 200–500 nM each in the reaction (optimize empirically).
• Annealing temperature: Optimize around predicted Tm; gradient qPCR can rapidly identify the optimal Ta.
• Reporting: Follow MIQE guidelines for primer disclosure and assay validation (sequence/amplicon info to be taken from CoA if provided).

Reaction Conditions

Item-specific status

• Validated reaction conditions are not provided for this product; refer to CoA/Spec Sheet.

General qPCR conditions (literature guidance; adjust to polymerase and instrument)

• Master mix: 1× qPCR mastermix containing hot-start DNA polymerase, dNTPs, Mg2+, and buffer. Choose dye-based (e.g., SYBR Green) or probe-based mix per assay design.
• Primer concentration: Start at 200–500 nM each; optimize to minimize primer–dimer while maintaining efficiency.
• Template: cDNA corresponding to 1–100 ng total RNA input equivalent per 20 µL reaction (optimize by sample type).
• Cycling (example): 95 °C 2–3 min (enzyme activation); 40 cycles of 95 °C 10–15 s (denaturation), 58–62 °C 20–30 s (annealing/extension). Include melt curve for SYBR assays.
• Amplicon length: Typically 70–200 bp for qPCR (actual for this item not specified).
• Efficiency/linearity: Evaluate via 10-fold serial dilutions; target 90–110% efficiency, R2 ≥ 0.99.
• Additives: For GC-rich targets, consider 2–5% DMSO or 0.5–1 M betaine; titrate carefully and validate specificity.

Controls and QC

• NTC, no-RT, and positive control RNA/cDNA.
• Specificity checks: Single melt-curve peak and clean single band on gel (optional). Sequence confirmation of the amplicon is recommended during initial validation.

Safety and Handling

Item-specific safety (from Product Data)

• GHS classification, signal word, pictograms, H-statements: Not specified for this item; refer to SDS.
• Research use: For research use only.

General safety guidance for DNA oligonucleotide primers (literature/best practice)

• Expected hazards: DNA oligos are generally considered of low toxicity; however, handle as laboratory chemicals. Avoid ingestion, inhalation of powders/aerosols, and contact with eyes/skin.
• PPE: Laboratory coat, gloves (nitrile), and safety glasses. Change gloves frequently to prevent nuclease contamination.
• Nuclease control: Work in clean, nuclease-free area. Use filtered tips and nuclease-free tubes/water. Wipe benches with DNA/RNase decontamination solutions if needed.
• Incompatibilities: Avoid strong acids/bases and oxidizers that degrade nucleic acids. Avoid metal contamination that can catalyze degradation.
• First aid overview: If inhaled (aerosols), move to fresh air; if on skin/eyes, rinse with water for several minutes; if ingested, rinse mouth and seek medical advice. Always consult SDS for authoritative instructions.
• Special risks: Repeated freeze–thaw cycles can shear/degrade oligos and increase nuclease risk in shared freezers; aliquot upon first thaw. Maintain cold chain for any supplied aqueous formulations.
• Waste: Dispose of small volumes with nonhazardous aqueous waste per institutional policy; decontaminate DNA-containing waste when required by protocol.

Solvent Selection

Item-specific guidance (from Product Data)

• No solvent is specified with the product. Storage at −20 °C and avoidance of repeated freeze–thaw are indicated.

General recommendations for DNA primer dissolution (literature/best practice)

• Primary solvents: Use nuclease-free water for immediate use. For longer-term stability, reconstitute in low-TE buffer (e.g., 10 mM Tris-HCl, 0.1–1 mM EDTA, pH 7.5–8.0) to chelate divalents and suppress nuclease activity.
• Concentration targets: Prepare 100 µM stocks (common) and 10 µM working solutions. Filtered, sterile, nuclease-free solutions are essential.
• Miscibility/pH: Aqueous buffers only; avoid organic solvents which can denature polymerases or alter primer behavior. Maintain near-neutral pH.
• Salt considerations: High Na+ stabilizes duplexes; however, final qPCR buffer composition is set by the master mix. Keep primer stocks in low-salt solutions to avoid precipitation on freezing.

When to choose alternatives

• If primer solubility is poor at very high concentrations, gently warm to 37 °C and vortex briefly. Do not use DMSO for primer stock unless protocol explicitly requires it; introduce any enhancers (e.g., DMSO, betaine) at the reaction mix stage, not in the stock solution.

Small comparison (literature)

• Nuclease-free water: Maximum flexibility; slightly less long-term stability.
• Low-TE: Improved long-term stability; EDTA can chelate Mg2+, so maintain adequate dilution before adding to Mg2+-containing PCR mixes.

Storage and Reconstitution

Item-specific instructions (from Product Data)

• Storage: Store at −20 °C. Avoid repeated freezing and thawing.
• Shipping: Shipped in ice chest with ice pads.

General best practices (literature)

• Upon receipt: If lyophilized, centrifuge briefly before opening to collect material. Reconstitute each primer in nuclease-free water or low-TE buffer to a convenient stock concentration (e.g., 100 µM). Mix gently and allow 5–10 min for complete dissolution.
• Aliquoting: Prepare single-use aliquots (e.g., 10–50 µL) to prevent multiple freeze–thaw cycles. Label with concentration, buffer, and date.
• Storage conditions: Store aliquots at −20 °C protected from light. For long-term storage (>12 months) of lyophilized material, keep desiccated and at −20 °C. Avoid frost-free freezers that cycle temperature.
• Stability considerations: EDTA in low-TE buffers chelates Mg2+ to suppress nuclease activity, improving stability. Avoid repeated thawing; keep on ice during setup.
• Working solutions: Keep 10 µM working stocks at 4 °C for up to 1–2 weeks (laboratory practice) and at −20 °C for longer; verify performance periodically.

Documentation

• Record lot number, reconstitution buffer, concentration (by A260 if possible), and storage location. For critical assays, confirm concentration via UV spectrophotometry using sequence-specific extinction coefficients (if provided on CoA).

Structure and Identity

Item-specific facts (from Product Data)

• SKU: H746972
• Product name: Human PPIH qPCR Primer Pair
• Type: qPCR primer pair (DNA oligonucleotides), designed to amplify human PPIH transcript(s)
• CAS, CID, InChIKey, SMILES: Not applicable to this oligonucleotide mixture; not specified for this item; refer to CoA/Spec Sheet.

General description (literature)

• Composition: Two single-stranded DNA oligonucleotides (forward and reverse primers), typically 18–30 nt each, lyophilized or supplied in aqueous buffer.
• Structural features: Linear, phosphodiester-linked deoxyribonucleotides with 5′-OH and 3′-OH termini (unless otherwise modified; modifications not specified for this item). No stereocenters beyond the native D-deoxyribose chiral centers common to DNA.
• 2D structure in words: Repeating deoxyribose–phosphate backbone with pendant nucleobases (A, T, G, C) projecting from the sugar; the two primers are non-complementary to each other (ideally) and complementary to opposite strands of the PPIH target locus.

Notes and identifiers

• Molecular formula / molecular weight: Sequence-dependent and not specified for this item; refer to CoA/Spec Sheet.
• Target gene: PPIH (Peptidylprolyl isomerase H), species: Human (as indicated by product name).

Synthetic Utility

Applicability

• As a pair of DNA primers, this product is not a synthetic building block or reagent in classical organic synthesis.

Relevant laboratory utility (general/literature)

• Molecular cloning support: Primers may be repurposed for endpoint PCR to generate amplicons for cloning (e.g., TA cloning) if design parameters permit; sequence/amplicon features for this item are not specified.
• Quantitative assay development: Useful in developing standard curves, assessing assay linearity, and validating reference gene panels in transcriptomics workflows.
• Method validation: Assists in optimizing polymerase choice, Mg2+ concentration, and annealing temperatures in qPCR platforms.
• Compatibility: Generally compatible with SYBR Green and hydrolysis-probe chemistries (primers are common to both); the latter requires a separate probe (not part of this product).

Practical notes

• Without specified sequences, users should consult the CoA for GC content, predicted Tm, and amplicon length to plan annealing/extension conditions and confirm specificity by BLAST.
• For cloning or sequencing applications, ensure primers include required overhangs/restriction sites if needed; no such features are specified for this item.

Target Specificity

Item-specific facts (from Product Data)

• Target: Human PPIH (Peptidylprolyl isomerase H) gene.
• Format: qPCR primer pair (forward and reverse primers) for amplification of human PPIH transcripts.

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

• Primer sequences (5′→3′)
• Amplicon size and genomic/exonic location
• Melting temperatures (Tm) and optimal annealing temperature
• Validated sample types and species cross-reactivity (if any)

General guidance (literature)

• Specificity considerations: Primers should uniquely match PPIH transcripts and avoid known pseudogenes or homologous cyclophilin family members. Intronic spanning or exon–exon junction targeting reduces genomic DNA amplification.
• Cross-reactivity risk: Cyclophilin family sequence similarity necessitates in silico BLAST verification against the human genome/transcriptome and, if applicable, evaluation in non-human samples.

Häufig gestellte Fragen

How should this product be stored?

Store at ?20 °C. Avoid repeated freeze–thaw cycles. Aliquot into single-use volumes before freezing.

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.