Twenty six transcripts had been detected from just one or two with the three samples. The signal intensities for these transcripts were low. Only one transcript in one sample had its signal intensity one,000, indicating the inconsistence between the duplicates was on account of very low signals of those transcripts. For the two 100 cell samples from MCF 7, 615 and 614 gene transcripts have been detected, respec tively, with 597 detected in each. Of these 597 transcripts, 562 had signal intensities differing much less than two fold. Similar to the scenario with NCI ADR RES samples, all 34 transcripts that have been detected in only one sample but not the other had low signal intensities with only nine genes whose signal intensities had been 1,000 in one within the two samples. Mainly because samples prepared from a substantial amount of cells are usually linked with high dependability, we even further com pared the microarray final results from the NCI ADR RES 100 cell samples with those from a 10,000 cell sample in the very same cell line.
Resulting data also present a large degree of corre lation. As proven in Table one, 630 within the 650 gene transcripts detected from all of the one hundred cell samples have been also detected through the 10,000 cell sample. Sixty 3 gene transcripts had been detected in no less than one on the 3 a hundred cell samples but not while in the 10,000 cell sample, or vice versa. Of these 63 explanation gene tran scripts, 61 had signal intensities beneath 1,000 in each of the three 100 cells. Nevertheless, the adjust from 100 to 10,000 cells did improve the detection of 21 gene transcripts whose signal intensities had been 2 fold greater from the 10,000 cell sample than people while in the 100 cell samples. Amongst these 21 transcripts, 6 had signal intensities within the 10,000 cell sample selleck a lot more than 15 fold better than the common intensities from the corresponding genes during the three a hundred cell samples, indicating that implementing 10,000 cells may have substantially increased the copy numbers of these transcripts or modified their absence status to presence.
These data indicate that our program not merely can make very trustworthy success even with as number of as a hundred cells but in addition is extremely delicate to your copy amount transform for your reduced copy variety gene transcripts. Sensitivity within the higher throughput gene expression profiling strategy To even further test the sensitivity of our substantial throughput gene expression profiling technique, single NCI ADR RES cell samples have been ready and applied for multiplex gene expression assay of your 1,135 mRNA species. Microarray final results from 3 of these samples are listed in Further file 3. The numbers of gene transcripts detected from your 3 single cell samples have been 590, 576, and 614, respec tively. Of those transcripts, 503 were detected from all sin gle cells. In the 503, 463 were also detected from all non single cell samples, indicating a prevalent expression of those genes in many, if not all, cells at reasonably high ranges.