Gene expression analysis by quantitative reverse transcription PCR (qRT–PCR) allows accurate quantifications of messenger RNA (mRNA) levels over different samples. Create two targets with the same SYBR DYE; Name your GENE (red) and REFERENCE gene (Green: GAPDH or Beta Actin) Use 2 (two) separate reaction tubes with the same sample name for […]
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How to check the performance of the thermal gradient block?
Real time PCR instruments has two main functions; thermal block and optical module, new generation peltiers control the heating and cooling sensitivity, LED excitation source and new generation CCD cameras are stated in the optical module. So, to check the performance of a RT-PCR; you have to check the performance […]
Read MoreWhat could be the reason of poor RT-PCR results?
Achieving accurate and reliable results depends on: diagnostic kit, RNA prep solution, swap for sampling, disposables (pipette tips, strip tubes and plates for RT-PCR) and instrument. Possible Reasons of Poor Results; Not to store IVD kit at -20 C in a deep freezer (Please check the expiration date) To use […]
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Read MoreRT-PCR Result: Lower Amplification
Low amplification plots may be caused by; Low quality PCR strip tubes, optical tubes, plates or sealing may be used Master mix may be melted many times (more than 3 times), efficiency may be down Kits may be out of date, please check the expiration dates of the RNA isolation […]
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