Description

[Product Specifications]

1. TL50 50*3 Reactions
2. TL100 100*3 Reactions

[Product Description] Telomeres are non-coding tandem repeat (TTAGGG)n arrays at the ends of linear chromosomes in eukaryotes. Changes in telomere length are associated with aging, cancer, neurodegenerative diseases, and other conditions. Quantitative PCR is a classic technique for telomere length detection, featuring short testing time, simple operation, accurate results, and suitability for batch sample testing.

This kit uses dual-color fluorescence qPCR (SYBRgreen+VIC) to detect relative telomere length in DNA from blood, saliva, or cells. The kit features the following characteristics:

1. Stable: Closed-tube operation prevents cross-contamination.
2. Reliable: Single-tube detection of telomeres and internal reference reduces well-to-well interference. Multi-copy internal reference gene reduces quantity difference with telomeres.
3. Convenient: Simple operation, no electrophoresis required.
4. Quantitative: ΔCT values enable quantitative detection of relative telomere length.

[Transportation and Storage Conditions] Shipped frozen at low temperature, store at -20°C, shelf life 1 year. After thawing, store at 4°C for up to 4 weeks.

[Kit Components] [Table with components]

Note: Before using the provided reagents, gently invert ten times to mix, then centrifuge at low speed and open carefully. Avoid bubbles. During experimentation, operate on ice.

[Operating Procedures]

1. Sample Quality Control Extract DNA from blood, cells, or tissue using commercial DNA extraction kit. DNA purity A260/A280≈1.8, standardize sample concentration to 1520ng/µL, load 4µL. Note: Standards are pre-standardized to 1520ng/µL, load 4µL. Excessively high or low concentrations of samples and standards affect amplification efficiency, interfering with experiments and increasing measurement error.
2. qPCR Reaction System and Conditions 20µL reaction system example [Reaction system table]

PCR instrument settings using ABI 7500 as example. Other quantitative PCR instruments should calibrate parameters using standards or consult technical staff.

[Baseline Settings] Internal reference baseline: Set using standard baseline, set to 612 cycles. Telomere baseline: Set using standard baseline, set to 37 cycles.

[Threshold Settings] Internal reference threshold: Set using standard CT values, set to 17±1. Telomere threshold: Set using standard CT values, set to 12±1.

Notes: (1) If standard internal reference CT value >25 but negative control internal reference CT value normal, indicates kit amplification efficiency decrease or instrument malfunction. (2) If negative control internal reference or telomere CT<25, indicates minor contamination in environment or sample loading process.

Remarks:

1. Sample testing and standards should have three technical replicates, negative control single replicate.
2. For blood DNA samples, obtained relative telomere length can be compared with Shanghai Biowing Applied Biotechnology Co., Ltd.’s 30,000+ Chinese population database to evaluate individual telomere relative length; for other samples, recommend establishing own database for further analysis.
3. Telomere length calculation method (T/S): First calculate ΔCT=CT(telomere)-CT(internal reference) for standards and samples Then calculate T/S=2-(ΔCT(sample)-ΔCT(standard)) Note: T represents test sample, S represents standard, T/S represents relative telomere length ratio between test sample and standard.