All products have special prices for bulk purchase, please contact for more details if required.

Cat. No.: HTASTM-50 (for 50T)

Cat. No.: HTASTM-200 (for 200T)

Description

Human Telomerase Activity Assay Kit by SYBR Green qPCR for TERT mRNA—also referred to as the Human Telomerase Reverse Transcriptase Subunit Assay Kit by qPCR with SYBR Green or the hTERT qPCR Assay Kit—is designed to rapidly and sensitively measure the expression level of the telomerase catalytic subunit (hTERT) in human cells or tissues using specific primers targeting the TERT gene and the internal control gene GAPDH. The hTERT mRNA level serves as an indirect indicator of telomerase activity. This kit is a contamination‑preventive qPCR assay that contains high‑quality UDG enzyme and dUTP in optimized proportions to effectively eliminate false positives or artificially low CT values caused by carryover contamination during PCR amplification.

Telomerase is a specialized reverse transcriptase composed of protein and RNA, directly involved in the synthesis of telomeres at the ends of eukaryotic chromosomes. In normal somatic cells, telomeres gradually shorten with each cell division, whereas increased telomerase activity can effectively maintain telomere length. Abnormally elevated telomerase activity can lead to uncontrolled cell proliferation and tumorigenesis. Except for normal human leukocytes (such as activated B and T lymphocytes), human germline tissues (adult testes and ovaries, excluding mature sperm and oocytes), and proliferating stem cells, telomerase activity is generally undetectable in normal somatic cells. However, most human cancers, as well as certain precancerous lesions and benign tumors, exhibit abnormally high telomerase activity. Therefore, highly sensitive and rapid detection of telomerase activity is of great significance for early cancer diagnosis.

Human telomerase consists of three major components: human telomerase reverse transcriptase (hTERT), human telomerase RNA (hTER or hTR), and human telomerase‑associated proteins. Among these, hTERT is considered the key determinant of telomerase activity. The mRNA level of hTERT correlates closely with overall telomerase activity, making hTERT mRNA a reliable indicator for assessing telomerase activity.

Currently, the most commonly used methods for detecting telomerase activity fall into direct and indirect approaches.

• Direct methods detect whether telomerase can extend a DNA template, and evaluate extension length through nucleic acid electrophoresis or probe hybridization. Examples include the Telomerase Repeat Amplification Protocol (TRAP), telomere repeat extension assays, colorimetric assays, fluorescence assays, and surface‑enhanced Raman spectroscopy. These methods typically require protein extraction, involve complex procedures, have limited sensitivity, or require radioactive labeling (posing safety and environmental challenges), or suffer from poor reproducibility and long turnaround times.

• Indirect methods measure the mRNA level of the telomerase catalytic subunit hTERT as a proxy for telomerase activity. This approach is simpler, requiring only mRNA extraction, reverse transcription to cDNA, and qPCR, while offering high sensitivity and accuracy. This kit adopts the indirect detection method.

This kit uses mRNA from the telomerase TERT gene and the internal control GAPDH gene as detection targets. It provides optimized reverse transcription primers (Specific RT Primers (10X), TERT Primer Mix (10X), and GAPDH Primer Mix (10X)), along with the required qPCR premix SYBR Green qPCR Mix (2X, UDG). The premix contains BeyoFast™ Taq DNA Polymerase, UDG enzyme, PCR buffer, dNTPs, dUTP, SYBR Green I dye, stabilizers, and magnesium ions—everything needed for qPCR. In addition, the kit includes a Positive Control to verify that the kit is functioning properly.

UDG (Uracil-DNA Glycosylase), also known as UNG (Uracil-N-glycosylase), catalyzes the hydrolysis of the N‑glycosidic bond between uracil (dU) bases and deoxyribose in uracil‑containing DNA strands, thereby releasing free uracil. It is primarily used to eliminate carryover contamination from previous PCR amplifications.

The contamination‑prevention mechanism works as follows: an appropriate amount of dUTP is added to the PCR reaction so that dUTP replaces dTTP during DNA synthesis, generating PCR products containing dU bases. In subsequent PCR reactions, UDG selectively cleaves any single‑stranded or double‑stranded DNA containing dU that may have been introduced as contamination from earlier PCR products, thereby preventing false positives or interference caused by carryover amplicons.

This kit provides both Low ROX and High ROX formulations, making it broadly compatible with qPCR instruments that require no ROX, Low ROX, or High ROX as a passive reference dye. ROX is used to correct fluorescence fluctuations unrelated to PCR amplification, minimizing well‑to‑well variation. Such variation may arise from factors like pipetting errors or sample evaporation.

Different qPCR instruments have different ROX requirements. Please select High ROX, Low ROX, or no ROX according to the specifications of your instrument when preparing the reaction mixture.

Components

• SYBR Green qPCR Mix (2X, UDG)
• Specific RT Primers (10X)
• TERT Primer Mix (10X)
• Positive Control (10X)
• Low ROX (50X)
• High ROX (50X)
• Ultrapure Water