Isolated RNA that met the quantity and quality specifications from each of the slide mounted sections was tested twice in separate assay runs.
For high-throughput RNA extraction, we found that for most tissues, the SimplyRNA HT kit was able to isolate RNA that meets the quantity and quality criteria for downstream analysis via qPCR, microarray and NGS technologies.
Likewise, the SimplyRNA HT kit extracted RNA meeting the quantity and quality criteria for qPCR analysis from spleen, kidney and larvae; though in our hands, the ability of this kit to extract suitable RNA from blood samples was less consistent.
An extraction was considered successful if it met the quantity and quality set by the 1KP consortium agreement: ≥30 μg of total RNA, RNA integrity number (RIN) higher than 5, and rRNA ratio (26S/18S) greater than 1.
To meet the sample quantity for SOLiD sequencing, the complementary DNA libraries were then further amplified using 15 cycles of PCR.
However, this kit was less consistent in its ability to extract RNA from blood and spleen as the majority of the blood and spleen RNA samples extracted with this kit did not meet the minimum quantity requirements for microarray analysis.
In order to meet the sample quantity for SOLiD sequencing, the cDNA libraries were then further amplified using supplied primer sets containing different barcodes by 15 or 18 cycles of polymerase chain reaction.
However, the reality is that such "puny" orders cannot meet the MOQ (Minimum Order Quantity) for many components, resulting in a much higher BOM (bill of materials) than projected.
The relationship with availability is consistent with the finding of [29] who found that the main barrier to gas use in some villages in Pakistan was the lack of availability of the LPG in sufficient quantity to meet the need of the consumers.
A power system is designed to generate electric power in sufficient quantity to meet the present and estimated future demands of the users in a particular area, to transmit it to the areas where it will be used, and then distribute it within that area on a continuous basis.
