Sample Preparation
Use standard industry tools and methods to extract RNA from cell or cells in sample, purify the material and measure quality. If converting to cDNA use normal Reverse Transcription protocol.
ZS Genetics Labeling
ZSG labels are innovative, but the labeling process is not. The target sample, mRNA or cDNA, is separated into single strands using common PCR and polymerase enzymes that incorporate modified dNPTs from ZS Genetics (with its atomic labels). This step is repeated if different labels are needed for complex applications.
The application determines the number of bases labeled and the type of label used. If a scientist just wants to see molecules, say to count them for Gene Expression, then only 1 or two bases need to be labeled (probably Cytosine and Thymine) and the same label type can be used. This can be achieved in one PCR cycle. If more bases need to be labeled and the type of base matters, say for Sequencing, then a series of label incorporation steps is needed. There are 8 possible base positions (A, T, C & G on each cDNA double-strand), with 5 of them needing to be labeled in order to read the sequence based upon the label type (iodine, bromine, etc.) and position. Each label type must be separately identifiable in the TEM image, for example by “large” black dots, smaller black dots and large grey dots.
Substrate
The ZSG substrates are produced with semiconductor fabrication techniques. They are essentially micron-scale glass slides. They will be functionalized either as hybridization arrays or with non-sequence-specific linking agents.