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• Standard laboratory equipment
• Pipettes and nuclease free, aerosol-preventative tips
• Phosphate Buffered Saline (PBS) for washing the cells
• Protector RNase Inhibitor:
• Protector RNase Inhibitor 2,000 U; Cat. No. 03335399001
• Protector RNase Inhibitor 10,000 U; Cat. No. 03335402001

", "Country": "XG", "Code": "Additional Equipment and Reagent Required", "Name": "Additional Equipment and Reagent Required" }, { "Language": "en", "Value": "Easy-to-use reagent for cell lysis, prior to real-time RT-PCR applications.", "Country": "XG", "Code": "Product Characteristics", "Name": "Product Characteristics" }, { "Language": "en", "Value": "

Vial / Bottle	Function / Description	Content
1	RealTime ready Cell Lysis Buffer	200 ml easy-to-use Lysis Buffer

", "Country": "XG", "Code": "Content", "Name": "Content" }, { "Language": "en", "Value": "To obtain optimal results in downstream procedures, especially in real-time RT-PCR assays on the LightCycler^® Instruments, do not process samples larger than this reagent is designed to handle.

The optimal amount of sample material is as follows:

• For adherent cells only: Seed an appropriate number of adherent cells and grow them overnight.
• For suspension cells only: Cells can be lysed directly in their cell culture media, or first pelleted and optionally resuspended in PBS before lysis.
• The maximal cell number and the volume of RealTime ready Cell Lysis Buffer depends on the type of culture plate used, but one lysis reaction can be done on a range of 3 to 3 × 10⁴ cells.

Tissue Culture Plate Type	Number of Cells per Lysis Reaction (or per well)
96-well	Up to 30,000(1)
384-well	Up to 5,000
1536-well	Up to 2,000

1. Depending on the cell size, up to 10⁵ cells can be used. Although, higher concentrations may result in loss of signal, as the maximal number of adherent cells is restricted by cell density.

", "Country": "XG", "Code": "Sample Materials", "Name": "Sample Materials" }, { "Language": "en", "Value": "The RealTime ready Cell Lysis Buffer is designed for the lysis of cultured cells. Cell lysates can then be directly used for RT-qPCR or cDNA synthesis followed by a qPCR step. The quality of RNA isolated is suitable for relative quantification of mRNA by qPCR. The RealTime ready Cell Lysis Buffer is designed for rapid, high-performance gene expression studies using cultured cells in RT-qPCR or qPCR applications. The buffer uses an advanced one-step protocol for lysing 3 to 30,000 cells within 5 minutes at room temperature. The resulting lysate can be used directly for downstream real-time PCR applications, eliminating the time-consuming RNA purification step and ensuring minimal loss of RNA. With this large pack size and easy-to-use protocol, the RealTime ready Cell Lysis Buffer can be easily adapted to high throughput experiments while minimizing waste materials. In addition, this buffer easily integrates into automated cell-based gene expression workflows.", "Country": "XG", "Code": "Applications", "Name": "Applications" }, { "Language": "en", "Value": "RealTime ready Cell Lysis Buffer is an easy-to-use reagent for the lysis of cultured cells prior to reverse transcription. The buffer is designed for 5,000 lysis reactions, with a final volume of 40 μl each, or for 50,000 lysis reactions, with a final volume of 4 μl each.", "Country": "XG", "Code": "Intended Use", "Name": "Intended Use" }, { "Language": "en", "Value": "RealTime ready Cell Lysis Buffer is designed for the lysis of cultured cells. Cell lysates can be directly used for cDNA synthesis and subsequent qPCR on the LightCycler^® Instruments or PCR on standard thermal block cyclers.
The quality of RNA isolated is suitable for relative quantification of mRNA by Transcriptor One-Step RT-PCR Kit, especially on the LightCycler^® Instruments.", "Country": "XG", "Code": "Background Information", "Name": "Background Information" }, { "Language": "en", "Value": "For lot-specific certificates of analysis, see section Contact and Support.", "Country": "XG", "Code": "Quality Control", "Name": "Quality Control" }, { "Language": "en", "Value": "RealTime ready Cell Lysis Buffer is an easy-to-use reagent for lysing cultured cells in 96-/ 384-/1536-well plates, prior to reverse transcription or qPCR. This product is designed specifically for gene expression assays using a LightCycler^® Master Mix on the LightCycler^® 96 System (96-well format), LightCycler^® 480 System (96- or 384-well format), or the LightCycler^® 1536 System (1536-well format).

The isolation of RNA from experimental samples is the first step in gene expression studies. RNA isolation can be a quite time-consuming and intensive process, especially when dealing with samples with small amounts of material/cells or when dealing with a high number of samples. The RealTime ready Cell Lysis Buffer is designed to quickly lyse cultured cells prior to reverse transcription, without purifying the RNA, thus ensuring minimal loss of RNA and simplification of the whole gene expression workflow.", "Country": "XG", "Code": "Principle", "Name": "Principle" }, { "Language": "en", "Value": "RealTime ready Cell Lysis Buffer is an easy-to-use reagent for the lysis of cultured cells prior to reverse transcription.", "Country": "XG", "Code": "Product Description", "Name": "Product Description" }, { "Language": "en", "Value": "Total Time for Cell Lysis: 5 min
Optional wash step adds 5 min to the lysis protocol.", "Country": "XG", "Code": "Assay Time", "Name": "Assay Time" } ] } } ] }