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

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

Description

Human Telomere Length SYBR Green qPCR Kit, also known as the Human Telomere Length qPCR Kit with SYBR Green, is a kit designed to quantitatively measure the relative length of telomeres in human samples quickly, efficiently, and sensitively through dye-based qPCR (Quantitative PCR) amplification of telomere and reference gene DNA. This product is contamination-resistant, containing high-quality UDG enzyme and dUTP in optimized proportions, effectively eliminating false positives or low CT values caused by product contamination during PCR amplification.

Telomeres are protective complexes at the ends of eukaryotic chromosomes composed of short tandemly repeated DNA sequences (TTAGGG) and associated proteins. As cells divide, telomere length shortens. Cells with very short telomeres enter a state of senescence, an irreversible halt in cell growth. Therefore, telomere length serves as an important marker for determining whether cells have entered a state of senescence. Telomeres can also act as biomarkers of cumulative exposure to environmental and life experiences and are risk factors for major diseases. Increasing evidence suggests that telomere shortening negatively impacts health and is associated with numerous health problems, including aging and cancer. Thus, quantifying telomere length is crucial in many aspects of cell biology research, including chromosome instability, DNA repair, aging, apoptosis, cell dysfunction, and carcinogenesis.

This kit provides the necessary premix for qPCR experiments, Fast SYBR Green qPCR Mix (2X, UDG), and specific primers. The premix contains Fast Taq DNA Polymerase, UDG enzyme, PCR Buffer, dNTPs, dUTP, SYBR Green I fluorescent dye, stabilizers, and magnesium ions. The kit includes two pairs of primers: a Telomere Primer Mix based on telomere repeat sequences and a Control Primer Mix based on a single-copy gene (Acidic ribosomal phosphoprotein, PO) on chromosome 12, used for data normalization.

This kit is highly sensitive and has high amplification efficiency. The primers detect genomic DNA samples in the range of 1.25-20 ng/μl with an amplification efficiency of up to 95%. Genomic DNA from HeLa cells at different concentration gradients was amplified using the Telomere Primer Mix and Control Primer Mix to generate standard curves. The results showed that both standard curves exhibited good linearity, indicating stable amplification performance and high accuracy in quantifying relative telomere length.

This kit provides Low ROX and High ROX, making it widely compatible with fluorescence quantitative PCR instruments that either require no ROX or need Low ROX or High ROX as calibration dyes. The role of ROX is to correct fluorescence fluctuations unrelated to PCR, thereby minimizing inter-well variability. Such variability may arise from various factors like pipetting errors and sample evaporation. Different fluorescence quantitative PCR instruments have varying requirements for ROX; please choose High ROX, Low ROX, or no ROX according to your specific instrument when preparing the reaction system.

Components

• Fast SYBR Green qPCR Mix (2X, UDG)
• Telomere Primer Mix (20X)
• Control Primer Mix (20X)
• Ultrapure Water
• Low ROX (50X)
• High ROX (50X)