BioReagent BioReagent for sensitive chromatographic and analytical workflows requiring minimal baseline interference.
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Cited in 0 peer-reviewed publications across chromatography, organic synthesis, and cross-coupling reactions.
Human GAPDH qPCR Primer Pair is primarily designed for SYBR Green‑based qPCR, one‑step qRT‑PCR, or semi‑quantitative PCR. This primer pair is pre‑designed, qPCR‑validated, and supplied as a premixed primer pair.
qPCR (Quantitative PCR), also known as real‑time fluorescent quantitative PCR or real‑time PCR, is a method that quantifies the total product accumulated after each polymerase chain reaction (PCR) cycle during DNA amplification using fluorescence detection. The two common approaches for qPCR are fluorescent dye‑based methods (e.g., SYBR Green) and probe‑based methods. SYBR Green‑based methods use a non‑specific fluorescent DNA‑binding dye (such as SYBR Green) to detect the accumulating PCR amplicons during the reaction. The probe method (often called the TaqMan probe method) does not use fluorescent dyes but instead employs DNA probes labelled with a fluorophore and a quencher that target the specific sequence to be detected by PCR.
For dye‑based methods like SYBR Green, the primers are critical. This series of primer products is developed using Aladdin’s proprietary primer design algorithm, with optimised sequences that have been validated to ensure good specificity, high amplification efficiency, low incidence of primer‑dimer formation, and reliable qPCR data. Most amplicons in this series are approximately 100–150 bp in length. This product is supplied as a pre‑mixed lyophilised powder. Each tube contains 1 nmol of forward primer and 1 nmol of reverse primer (total 2 nmol), is nuclease‑free, and can be reconstituted by adding 400 μL of ultrapure water to obtain a 2.5 μM each solution ready for use. When using 2 μL of primer mix per 20 μL or 25 μL reaction, each tube provides sufficient material for 200 qPCR reactions.
Gene Information
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Amplicon Information
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Precautions
1.The length of the PCR amplicon may vary due to the existence of multiple splicing isoforms after gene transcription.
2.Although this series of primer products has good specificity, we still recommend performing a melt curve analysis to confirm the specificity of the amplification reaction. If only a single melt curve peak (corresponding to the melting temperature, T<sub>m</sub>, of the double‑stranded DNA product) is observed, it indicates a single specific product. If the melt curve shows double peaks, multiple peaks, or irregular peaks, it may indicate primer dimers, non‑specific amplification, genomic DNA contamination, or contamination of reagents or the environment. We recommend including a no‑template control (NTC) – i.e., a reaction containing all components except the template – so that by comparing the melt curves of sample wells and the NTC well, the presence of primer dimers or other non‑specific amplification can be judged.
3.If amplicon contamination is present in the reaction system, we recommend using the contamination‑proof UltraBio™ SYBR Green qPCR Mix (S751589).
4.This product is for professional scientific research use only. It must not be used for clinical diagnosis or treatment, nor for food or drug applications, and should not be stored in ordinary domestic premises.
5.For your safety and health, please wear a lab coat and disposable gloves when handling.
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Notes:
1.Typically, a final primer concentration of 0.2–0.5 μM each gives good detection results; the concentration can be adjusted within the range of 0.1–1.0 μM each as needed.
2.The recommended amount of DNA template is generally 1–10 ng of cDNA. Because the copy number of the target gene varies among different species, you may increase the template amount or perform serial dilutions to determine the optimal template quantity. When using cDNA directly from an RT‑PCR reaction, its volume should not exceed 10% of the total PCR reaction volume.
3.The recommended reaction volume for 96‑well plates is 20 μL; you may scale the reaction up or down proportionally according to actual experimental needs.
4.It is recommended to include a no‑template negative control.
(4) Gently pipette or vortex briefly to mix, then centrifuge at room temperature for a few seconds to collect the liquid at the bottom of the tube.
(5) Place the prepared PCR reaction tubes in a real‑time PCR instrument and start the quantitative PCR reaction.
2. PCR reaction program
Prior to the real‑time PCR reaction, pre‑denature the template – typically set at 95 °C for 2 minutes; for complex or high‑GC templates, extend the time appropriately to 5–10 minutes.
(1) Pre‑denaturation: 95 °C for 2 minutes
(2) Denaturation: 95 °C for 15 seconds
(3) Annealing/Extension: 60 °C for 15–30 seconds
(4) Repeat steps (2) and (3) for a total of 40 cycles
(5) Melt curve analysis (optional): 95 °C for 15 seconds, 60 °C for 15 seconds, 95 °C for 15 seconds
(6) Analyse the results using the software provided with the real‑time PCR instrument.
Note: The above example is a conventional qPCR reaction system and is for reference only. Actual reaction conditions vary depending on the structure of the template, primers, etc. Optimal conditions should be determined according to the specific features of the template, primers, and target fragment, and the reaction system can be scaled up or down proportionally. The above is a two‑step qPCR; if a three‑step qPCR is preferred, simply add a 72 °C for 30 seconds step after the annealing/extension step, then repeat steps (2), (3) and this additional step for a total of 40 cycles.
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| Lot Number | Certificate Type | Date | Item |
|---|---|---|---|
 ZJ26F0737151 | Certificate of Analysis | Jul 17, 2026 | H746960 |
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