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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What 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: No Amplification
To check whether no amplification is caused by kir or instrument; Deselect the Subtract Baseline and Normalization functions If the lines are unexpected curves as above then it proves that instrument is collection data so you have to focus on your protocol or kits If the there is only a […]
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or sealing may be used Master mix may be melted many times (more than 3 times), efficiency may be down Kits may be out of date, please check the expiration dates of the RNA isolation […]
Read MoreRT-PCR Result: Normal Amplification
This is an expected Real Time PCR results because; (FAM: Specific regions for Covid-19 virus, HEX: Internal Control, RP gene for extraction) Positive control (PC) is amplified normally (This proves that PCR is done, instrument and kit is working properly) Negative Control (NC) has no amplification, so it proves that […]
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