Troubleshooting guide (Biomedicine T5, MolMed HT11) - Ping-Pong
Contents
1. Troubleshooting guide Biomedicine T5 MolMed HT11 Possible causes Probable solutions Very few cells in the culture thus no RNA amp poor FACS data No or very low CD93 mRNA expression One of the SNP assay did not work Wrong cell count due to the difficulty in distinguishing RBC from leukocytes Cells were washed away and lost Failed reverse transcription No cDNA Failed amplification on CD93 bad assay High apoptosis and low neutrophils population time frame Mixed cell population and different cell types have different CD93 mRNA expression levels Bad assay No DNA Bacterial contamination Not working in sterile lab Add RBC lysis buffer step Culture more cells Use 14mL tubes which gives firmer and protected pellets Check reference gene Check known positive controls Count live and dead cells when we harvest them and compare Cell type specific culture Grow more cells Run the products on an agarose gel Positive control from known samples Check the other assay it worked Count the cells to see Work in sterile area hood or similar Put the lid on as soon as possible Increase antibiotics concentration in the media Some indicators eg pH to monitor Working with a bacteria incubator Don t do that Colleagues extensive use of bacteria Clean it well or well culture than past years No or degraded RNA Presence of RNase Use RNA stablilizing agent or RNase inh2. ibitor at all times Not low enough temperature Long processing time Low intensity for CD93 Removal of jelly may have removed Work sterile in ELISA some cells and soluable CD93 Soluble CD93 stuck on bacteria Culture more cells 24h lifespan of neutrophils Decrease grow time faster handling time
Download Pdf Manuals
Related Search