Abstract
With this report we aim to make available a standard operating procedure (SOP) developed for RNA stabilization of small blood volumes collected via a finger stick. The anticipation that this procedure may be improved through peer-review and/or readers public comments is another element motivating the publication of this SOP. Procuring blood samples from human subjects can, among other uses, enable assessment of the immune status of an individual subject via the profiling of RNA abundance using technologies such as real time PCR, NanoString, microarrays or RNA-sequencing. It is often desirable to minimize blood volumes and employ methods that are the least invasive and can be practically implemented outside of clinical settings. Finger-stick blood samples are increasingly used for measurement of levels of pharmacological drugs and biological analytes. It is a simple and convenient procedure amenable for instance to field use or self-collection at home using a blood sample collection kit. Such methodologies should also enable the procurement of blood samples at high frequency for health or disease monitoring applications.
Highlights
Use of sample collection methods that are least invasive and that can be practically implemented outside of clinical settings, Finger-stick blood collection is used for a wide range of applications in routine clinical practice
It is for instance by this means that millions of individuals collect daily small blood volumes to monitor sugar levels
Blood transcriptome profiling has proven useful in generating high-resolution molecular phenotypes: to investigate pathogenesis of a wide range of diseases[8,9,10,11]; to develop biomarker signatures[12,13,14,15]; and to assess response to vaccines or therapies[7,16,17,18,19,20]
Summary
Authors: Discussion of volume is extensive and warranted as this is in our experience one of the main causes of failure during the extraction step; we provide a procedure detailing “preparation of the sample collection tubes” (Please see “Material and Methods”); the capillary needs to be filled in order to collect appropriate volumes Alternative methods, such as use of micropipettes is possible if preferred; other products are available and could be tested for that purpose. The paragraph above was included in the manuscript (Part of Introduction) This point was addressed in response to Dr Cliff’s comment: In Our initial experiment with RNA extraction of 50 uL blood collection protocol in Tempus solution, we obtained the average RNA yield was 1 ug, Purity of RNA; average of RNA integrity number (RIN) was ~7.5 and A260/A280 ratio of ~2.14. The benefits of publishing with F1000Research: Your article is published within days, with no editorial bias You can publish traditional articles, null/negative results, case reports, data notes and more The peer review process is transparent and collaborative Your article is indexed in PubMed after passing peer review Dedicated customer support at every stage
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