1. Product Overview

TruDeteck™ is a new cell line and human and mouse sample identification technology based on deep sequencing, aiming to solve the problems of cell line mislabeling and contamination. This technology provides comprehensive cell line authentication, contamination detection and multi-species cross-contamination identification services through high-precision single nucleotide polymorphism (SNP) analysis. TruDeteck™ is not only suitable for human cell lines, but also supports the identification of mouse cell lines and other common species, ensuring the accuracy and reliability of research.

2. Product features

2.1 High-precision cell line authentication

·  200 human SNP sites : can be used to identify more than 2,000 human tumor cell lines.

·  400 mouse SNP sites : can be used to identify more than 100 common mouse tumor cell lines.

·  100 sets of human-mouse homologous fragments : used to accurately quantify the percentage of human-mouse in mixed human-mouse samples (such as PDX models).

·  6 segments of human Y chromosome : used to determine the gender of human samples.

2.2 Pollution detection and quantification

·  Mycoplasma-specific sequence : can identify whether a sample is contaminated with mycoplasma and quantify the percentage of contamination.

·  Unique sequences of multiple viruses : Can identify whether a sample is infected by common viruses and quantify the percentage of contamination.

·  More than ten other species-specific sequences : can identify whether samples are contaminated by other common species (such as hamsters and rats).

2.3 High sensitivity and accuracy

·  Detection sensitivity : The detection sensitivity of the SNP method based on deep sequencing can reach <1%, which is much higher than the 5-20% of the traditional STR method.

·  Accurately identify impurity cell lines : Not only can impurity cell lines be identified, but the contamination ratio can also be quantified.

·  Dynamic monitoring of mixed cell samples : The component ratios of multiple mixed cell samples can be dynamically monitored to study the differential killing effects of drugs on different cells.

2.4 Rich functions

·  Establish a standard SNP fingerprint library : A standard SNP fingerprint library can be established for any human sample library and subsequent identification can be performed.

·  Calculate genetic similarity : The genetic similarity between different samples of the same cell line can be calculated and an evolutionary tree can be established for traceability analysis.

·  Determine genetic background : The genetic background of human samples can be determined, such as ethnic information.

·  Determine gender : The gender of human samples can be determined.

3. Technical principles

3.1 SNP method based on deep sequencing

·  SNP (Single Nucleotide Polymorphism) : A variation of a single base pair in the genome that has unique combinations in different cell lines.

·  Sequencing steps :

i.  DNA extraction : Extract DNA from cell lines .

ii.  PCR amplification : Amplify specific SNP sites through PCR.

iii.  Deep sequencing : Use NGS technology to detect and analyze specific SNP sites to obtain the base information and frequency of each site.

iv.  Data analysis : Compare the results with known cell line SNP fingerprints in the database to confirm cell line identity or detect contamination.

3.2 Comparison with traditional STR method

·  STR (short tandem repeats) :

oPrinciple  : Short, repetitive DNA sequences in the genome, STR combinations are unique to each cell line .

Step o  : Extract DNA, amplify specific STR sites through PCR, separate and detect the length of the amplified product by electrophoresis, and compare it with the STR spectrum of known cell lines in the database.

oLimitations  : STR polymorphisms increase the difficulty of analysis and affect the confirmation of cell lines; the detection sensitivity is low (5-20%).

·  SNP method :

oAdvantages  : Higher sensitivity and accuracy, can identify impurity cell lines and quantify the proportion of contamination; can calculate the genetic similarity between different samples of the same cell line and establish an evolutionary tree; can determine genetic background and gender .

4. Sample requirements and testing procedures

4.1 Sample requirements

·  Animal tissue : 50 mg tissue sample, try to remove fascial tissue and necrosis, and quickly freeze in liquid nitrogen.

·  Cells : 10^6-5*10^6 cells were centrifuged to remove the culture medium, rinsed twice with pre-cooled PBS, centrifuged to remove as much supernatant as possible, and quickly frozen in liquid nitrogen.

·  Whole blood : 500μL EDTA anticoagulated whole blood, short-term storage at 4℃ or long-term storage at -20℃.

·  FFPE samples : 10-15 FFPE wax blocks or 5μm FFPE wax rolls, stored at room temperature.

·  Oral swab : Use oral swab to wipe 10 times on the left and right sides of the mouth respectively, and store in 1ml PBS.

4.2 Testing process

1.  Nucleic acid extraction and quality inspection : Ensure that the gDNA concentration is ≥100ng/μL, and A260/A280≈1.8-2.0.

2.  Library construction and quality inspection : Using the automated library construction workstation, 96 samples can be operated at the same time.

3.  Sequencing : high-throughput sequencing, pE150, 0.6Gb.

4.  Data analysis and reporting : standardized data analysis, customized data analysis, and generation of detailed reports.

5. Application scenarios

·  Biological sample bank management : ensure the authenticity and quality of samples and reduce research bias.

·  Drug research and development : Improve the accuracy of drug screening and clinical trials, and accelerate the development of new drugs.

·  Basic research : Support research on disease mechanisms, cell biology research, etc. to ensure the reliability of data.

·  Personalized medicine : used for accurate identification of patient samples to support the formulation of personalized treatment plans.

6. Conclusion

TruDeteck™ provides a comprehensive solution for authentication and contamination detection of biological samples through its high precision, high sensitivity and multi-functionality. Whether it is a human cell line or a mouse cell line, TruDeteck™ can ensure the accuracy and reliability of research, helping the efficient conduct of scientific research and drug development.