Buffer of Cell Lysate for Reverse Transcription

The study of gene expression often needs RNA preparation followed by cDNA synthesis and PCR. However, difficulty in obtaining a large amount of cells for RNA preparation is the bottleneck in some of the studies, such as laser dissected samples, FACS sorted cells and the cultured cells in 96-wells. Cell number is limited (<2,000 cells), which restricts RNA preparation. Signosis’ CL(TM) cell lysis buffer allows preparation of cell lysate, which can be used for direct reverse transcription without RNA preparation. Cell lysate made from a few cells is good enough for reverse transcription of RNA to cDNA, and following PCR analysis or cDNA plate array assays.

A. Benefits:

Signosis’ CL(TM) Cell Lysis Buffer offers following benefits:

  • Simplified procedures - Reverse transcription can be performed in cell lysate without RNA preparation.

  • Lower cell numbers -A few cells are good enough to make cDNA for PCR.

B. Principle of the technology

The buffer is optimized for both cell lysate preparation and reverse transcription. cDNA synthesis can be directly made from RNA in the cell lysate without RNA pre-isolation.

Figure 1
Diagram

Figure 2. Comparison of cell lysate amplification kit from Signosis with the related kit from competitor. A: 2000 HeLa cells were lysed with cell lysis buffer from a competitor (C) and from Signosis cell lysis buffer (S). cDNAs were synthesized with Signosis cDNA synthesis kit and the endogenous gene (actin) amplified with different cycles using Signosis PCR amplification system. B: The cell lysate from different cell numbers were used for actin amplification with 30 cycles.

 

 Product Size Cat. Price
Buffer of Cell Lysate for Reverse Transcription 100 CL-0001 $159
  • Cell Lysate Preparation for Reverse Transcription (PDF)

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