Real Time PCR with High Efficiency

UKR
PHOS
Post-Doc

Overall

Performance

Ease-of-Optimization

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Company:

Analytik Jena

Product Name:

qTOWER

Catalog Number:

844-00502-3

We are interested in minimum residual disease also called MRD. We used the qTOWER for our studies on MRD and we gained 2 points for Ct values.

Experimental Design and Results Summary

Application

MRD study

Starting Material

Bone Marrow infiltrated with tumor cells

Protocol Overview

RNA stabilisation and extraction efficiency using the PAXgene blood RNA tubes. RNA is extracted from PB, BM stabilised in PAXgene blood RNA tubes using the PAXgene blood RNA kit according to the manufacturer’s instructions with a DNAse digestion step before elution of RNA from the column. RNA samples (1 µg) were subjected to reverse transcription using Reverse Transcriptase (Roche) according to the supplier’s instructions. cDNA products were amplified on a qTOWER in 20 µl of reaction mixture containing the SYBR Green I Master (Roche) and 50 mM of forward and reverse primers Each amplification cycle consisted of an initial step at 95°C (5 minutes), followed by 45 cycles of denaturation at 95°C (10 seconds), annealing at 60°C (10 seconds), and extension at 72°C (30 seconds).

Tips

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Results Summary

We tested the performence of the qTOWER by dilution of tumor cells in BM. We gain 2 points for Ct values and we were able to detect 1 tumor cell in 10^6 BM cells. Image shows intensity/cycles, all experiments were done in triplicate and each color is another dilution.

Features Summary

None

Additional Notes

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Summary

The Good

Performance

The Bad

Program not intuitive enough.

The Bottom Line

Very good product, can be adapted to any purpose.

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