The EZ-press Cell to cDNA Kit PLUS II offers a fast and easy method to produce cDNA from cells in culture without RNA isolation. It is a good substitution for the TRIzol method in quantitative analysis of gene expression.
Besides the powerful genome DNA degrading efficiency equally with the previous B0003 Kit, it also has markedly improved cDNA stability than the B0003. cDNAs produced by kit can be stored at -80°C for up to 3 months with no obvious change in Ct value.
Meanwhile, it contains an inert blue dye in the 4× Color CRT Mix of this Kit to avoid of pipetting errors. It can be used with the EZBioscience® Color SYBR Green qPCR Mix (A0012, A0012-R1, A0012-R2), which is supplemented with an inert red dye. Mixing cDNAs, primers and other components with the EZBioscience® Color SYBR Green qPCR Mix together in a qPCR reaction turns the solution into purple. This provides a visual aid when pipetting, and decreases the risk of pipetting errors during reaction setup. And the dyes do not affect the specificity or sensitivity of qPCR reactions. So, these two kinds of kits are recommended to use together to get optimal results.
Moreover, this kit offers several significant advantages compared with TRIzol method:
1. Easy to use. There are just two steps (cell lysis and reverse transcription), no RNA isolation is needed.
2. Fast. The whole experiment (from cells to cDNA) can be completed in less than 25 minutes.
3. Stable and highly reproducible. The total RNA is perfectly reserved throughout the experiment, enabling stable and highly reproducible results.
4. High sensitivity. The lower detection limit for each sample is only 1000 cells.
5. Genome DNA is removed effectively by the gDNA Remover contained in this kit. Moreover, the reagents used in this kit are safe and non-toxic, which is pleasant compared with the smelly and hazardous TRIzol reagent.
Experimental Procedure at a Glance
It contains the following steps to get the cDNAs from cells by this Kit:
Preparation of cells
Cell Lysis
Reverse Transcription
And details are illustrated in the Product Manual.
Representative experimental results
cDNA synthesized by EZBioscience® EZ-press Cell to cDNA Kit and TRIzol - reverse transcription method were evaluated by real-time qPCR with 6 pairs of primers (in HEK-293T cells). Highly consistent results between EZ-press Cell to cDNA Kit and TRIzol -RT method can be observed in the following figure, which indicate that EZ-press Cell to cDNA Kit can completely replace TRIzol-RT method in preparing cDNAs from cells.
