EZCt Cells-2-Ct Direct

Request QuoteCatalog Number: EZCt-100mlSize: 100 ml

EZCt Cells2Ct Direct qPCR Lysis Buffer

mRNA purification for gene profiling by qPCR is becoming routine for functional genomics and personalize medical studies. mRNA is sentive to RNase degradation during the many steps of isolation, making the process less reliable and time consuming. Isolating mRNA in highthroughput gene expression or targeting applications for qPCR is almost impractical.

Reagents such as Cells-to-Ct^* that can be used to lyse cells in wells, and the resulting lysate could be used directly in qPCR would become the first choice. It is a very expensive 2-reagent protocol.

Lifeome scientists have developed the EZCt Cells2Ct Direct Lysis Buffer to vastly improve the process at a much affordable price. Lipid Aided RNase Inhibition (LARI) undergoes conformational changes upon contact with certain lipid molecules released from lysed cell membranes, and structurally inhibit RNase activity upon cell lysis. The inhibition is long lasting with the presence of those lipid molecules, allowing room temperature lysis and RNA stability.

- LARI Technology;
- Single Buffer;
- Room Temperature (RT) storage;
- Lysates are stable for up to 2 weeks, RT!
- 1-2-3 Protocol: wash, lyse, ready for qPCR!
- Results are in!

^{*: TM of ThermoFisher or its affiliates.}

Sizes

Cat. No.	Volume	Applications^*

EZCt-1ml:	1 ml	20 Wells
EZCt-10ml:	10 ml	200 (2 96-well plates)
EZCt-100ml:	100 ml	2000 (10 96-well plates)

*Est. based on 50 ul per well with less than 2000 cells of 96-well plate.
Custom size is also available upon request.

Specifications

Lot number:	On label
Formulation:	Clear to slightly turbid solution of proprietary formulation
Preservative:	None
Sterility:	0.2 um membrane filtered and packaged aseptically
Storage:	Room Temperature, 3-month Shelf-life, Do NOT freeze.
Lysate:	Stable at RT 1-2 weeks, -20°C for longer time

Protocols:

96 Well Plate:

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.

Storage and stability

Upon receiving, store the product at RT (20-30°C), with a shelf-life of 3 months.

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