Single Cell Gene Expression

The study of gene expression often needs RNA preparation followed by cDNA synthesis and PCR, but most of the time, you don't want to waste a large amount of cells for RNA preparation. This is a particular problem for researchers using laser dissected samples, FACS sorted cells, cultured cells in 96-wells, and liquid biopsy. 

 

Signosis’ Direct cDNA cell lysis buffer allows researchers to prepare cell lysate from small samples, which can be used for direct reverse transcription without RNA preparation. Cell lysate made from even a few cells is good enough for reverse transcription of RNA to cDNA, which you can then use for PCR analysis or cDNA plate array assays.

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Signosis Direct cDNA cell lysis buffer. A: The indicated cells were lysed with Direct cDNA cell lysis buffer from Signosis and competitor respectively, and subjected to RT-PCR for ß-actin with 30 cycles. B: Testing for genomic DNA contamination. Lane1. ß-actin was amplified with 36 PCR cycles directly from cell lysate without reverse transcription (RT). Lane2. ß-actin was amplified with 36 PCR cycles directly from cDNA transcribed from cell lysate.

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​Comparison of PCR products amplified from lysates made with Signosis Direct cDNA cell lysis buffer or TRIzol without DNAse treatment.  By using intron-spanning primers, DNA amplified from genomic DNA and reverse-transcribed mRNA can be distinguished.  PCR amplification from RNA prepared with TRIzol contains a band that corresponds with the predicted length of the genomic DNA, as well as the predicted length of the mRNA. However, PCR amplification from cell  lysate prepared with Signosis Direct cDNA cell lysis buffer only contains a band that corresponds with the predicted length of the mRNA and not the genomic DNA contamination.