96 Well Plates:
1. Treat your cells per your protocol;
2. Brifely wash cells with PBS.
3. Add 50 ul of EZCt lysis buffer (DNase I optional, and not included), and pipet up and down 5 times, avoid excessive bubbling;
4. Incubate the lysis reaction for 5 min before using (1 ul) for qPCR.
5. The rest of the lysate could be stored at RT for up to 2 weeks, or -20°C for longer time.
Human Blood:
There are about 4,000-10,000 white blood cells per microliter of blood.
1. Add 1ul of whole blood to 100ul room temperature PBS in a microcentrifuge tube. Tap the tubes to mix contents;
2. Briefely spin the tubes using a benchtop centrifuge or your regular procedure for collecting cells;
4. Resuspend the cell pellet in 100ul PBS;
5. Take 1ul of the resuspeded cell and add to a fresh tube containing 50ul of EZCt lysis buffer (DNase I optional, and not included), and pipet up and down 5 times, avoid excessive bubbling;
6. Incubate the lysis reaction for 5 min before using (1 ul) for (one-step) qPCR.
7. The rest of the lysate could be stored at RT for up to 2 weeks, or -20°C for longer time.
Single cell qPCR:
Tested by our customers who used 5 ul lysis buffer per cell.
If the product is going to be used for applications requiring absolute asepsis, it's best to filter-sterilize the solution using a sterile and non-pyrogenic 0.2 um membrane before use.
Upon receiving, store the product at RT (20-30°C), with a shelf-life of 3 months.
