RNA extraction

RNA extraction is t he purificat ion of RNA from biological samples. This procedure is
complicat ed by t he ubiquit ous presence of ribonuclease enzymes in cells and t issues, which
can rapidly degrade RNA.[1] Several met hods are used in molecular biology t o isolat e RNA
from samples, t he most common of t hese is guanidinium t hiocyanat e-phenol-chloroform
ext ract ion.[2][3] The filt er paper based lysis and elut ion met hod feat ures high t hroughput
capacit y.[4]

RNA ext ract ion in liquid nit rogen, commonly using a mort ar and pest le (or specialized st eel
devices known as t issue pulverizers) is also useful in prevent ing ribonuclease act ivit y.

RNase contamination

The ext ract ion of RNA in molecular biology experiment s is great ly complicat ed by t he
presence of ubiquit ous and hardy RNases t hat degrade RNA samples. Cert ain RNases can be
ext remely hardy and inact ivat ing t hem is difficult compared t o neut ralizing DNases. In addit ion
t o t he cellular RNases t hat are released t here are several RNases t hat are present in t he
environment . RNases have evolved t o have many ext racellular funct ions in various
organisms.[5][6][7] For example, RNase 7, a member of t he RNase A superfamily, is secret ed by
human skin and serves as a pot ent ant ipat hogen defence.[8][9] For t hese secret ed RNases,
enzymat ic act ivit y may not even be necessary for t he RNase's exapt ed funct ion. For
example, immune RNases act by dest abilizing t he cell membranes of bact eria.[10][11]

To count er t his, equipment used for RNA ext ract ion is usually cleaned t horoughly, kept
separat e from common lab equipment and t reat ed wit h various harsh chemicals t hat dest roy
RNases. For t he same reason, experiment ers t ake special care not t o let t heir bare skin t ouch
t he equipment .
 See also

Column purificat ion

DNA ext ract ion

Et hanol precipit at ion

Phenol-chloroform ext ract ion

References

1. Peirson SN, Butler JN (2007). "RNA extraction from mammalian tissues" (https://archive.org/detail
s/circadianrhythms00rosa/page/315) . Circadian Rhythms. Methods Mol. Biol. Methods in
Molecular Biology. 362. pp. 315–27 (https://archive.org/details/circadianrhythms00rosa/page/31
5) . doi:10.1007/978-1-59745-257-1_22 (https://doi.org/10.1007%2F978-1-59745-257-1_22) .
ISBN 978-1-58829-417-3. PMID 17417019 (https://pubmed.ncbi.nlm.nih.gov/17417019) .

2. Chomczynski P, Sacchi N (2006). "The single-step method of RNA isolation by acid guanidinium
thiocyanate-phenol-chloroform extraction: twenty-something years on". Nat Protoc. 1 (2): 581–5.
doi:10.1038/nprot.2006.83 (https://doi.org/10.1038%2Fnprot.2006.83) . PMID 17406285 (https://
pubmed.ncbi.nlm.nih.gov/17406285) . S2CID 28653075 (https://api.semanticscholar.org/CorpusI
D:28653075) .

3. Bird IM (2005). "Extraction of RNA from cells and tissue". Methods Mol. Med. 108: 139–48.
doi:10.1385/1-59259-850-1:139 (https://doi.org/10.1385%2F1-59259-850-1%3A139) . ISBN 1-
59259-850-1. PMID 16028681 (https://pubmed.ncbi.nlm.nih.gov/16028681) .

4. FortiusBio High Throughput RNA Extraction Filter Paper Card (http://fortiusbio.com/RNA_Extraction

_Card_8.html)

5. Rossier, O.; Dao, J.; Cianciotto, N. P. (2009). "A type II secreted RNase of Legionella pneumophila
facilitates optimal intracellular infection of Hartmannella vermiformis" (https://www.ncbi.nlm.nih.go
v/pmc/articles/PMC2662391) . Microbiology. 155 (3): 882–890. doi:10.1099/mic.0.023218-0 (http
s://doi.org/10.1099%2Fmic.0.023218-0) . PMC 2662391 (https://www.ncbi.nlm.nih.gov/pmc/articl
es/PMC2662391) . PMID 19246759 (https://pubmed.ncbi.nlm.nih.gov/19246759) .

6. Luhtala, N.; Parker, R. (2010). "T2 Family ribonucleases: Ancient enzymes with diverse roles" (https://
www.ncbi.nlm.nih.gov/pmc/articles/PMC2888479) . Trends in Biochemical Sciences. 35 (5): 253–
259. doi:10.1016/j.tibs.2010.02.002 (https://doi.org/10.1016%2Fj.tibs.2010.02.002) .
PMC 2888479 (https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2888479) . PMID 20189811 (http
s://pubmed.ncbi.nlm.nih.gov/20189811) .
 7. Dyer, K. D.; Rosenberg, H. F. (2006). "The RNase a superfamily: Generation of diversity and innate
host defense" (https://zenodo.org/record/1232824) . Molecular Diversity. 10 (4): 585–597.
doi:10.1007/s11030-006-9028-2 (https://doi.org/10.1007%2Fs11030-006-9028-2) .
PMID 16969722 (https://pubmed.ncbi.nlm.nih.gov/16969722) . S2CID 20922592 (https://api.sem
anticscholar.org/CorpusID:20922592) .

8. Harder, J. (2002). "RNase 7, a Novel Innate Immune Defense Antimicrobial Protein of Healthy
Human Skin" (https://doi.org/10.1074%2Fjbc.M207587200) . Journal of Biological Chemistry. 277
(48): 46779–46784. doi:10.1074/jbc.M207587200 (https://doi.org/10.1074%2Fjbc.M207587200) .
PMID 12244054 (https://pubmed.ncbi.nlm.nih.gov/12244054) .

9. Köten, B.; Simanski, M.; Gläser, R.; Podschun, R.; Schröder, J. M.; Harder, J. R. (2009). "RNase 7
Contributes to the Cutaneous Defense against Enterococcus faecium" (https://www.ncbi.nlm.nih.go
v/pmc/articles/PMC2712763) . PLOS ONE. 4 (7): e6424. Bibcode:2009PLoSO...4.6424K (https://u
i.adsabs.harvard.edu/abs/2009PLoSO...4.6424K) . doi:10.1371/journal.pone.0006424 (https://do
i.org/10.1371%2Fjournal.pone.0006424) . PMC 2712763 (https://www.ncbi.nlm.nih.gov/pmc/articl
es/PMC2712763) . PMID 19641608 (https://pubmed.ncbi.nlm.nih.gov/19641608) .

10. Huang, Y. -C.; Lin, Y. -M.; Chang, T. -W.; Wu, S. -J.; Lee, Y. -S.; Chang, M. D. -T.; Chen, C.; Wu, S. -H.;
Liao, Y. -D. (2006). "The Flexible and Clustered Lysine Residues of Human Ribonuclease 7 Are Critical
for Membrane Permeability and Antimicrobial Activity" (https://doi.org/10.1074%2Fjbc.M6073212
00) . Journal of Biological Chemistry. 282 (7): 4626–4633. doi:10.1074/jbc.M607321200 (https://
doi.org/10.1074%2Fjbc.M607321200) . PMID 17150966 (https://pubmed.ncbi.nlm.nih.gov/1715
0966) .

11. Rosenberg, H. F. (2008). "RNase a ribonucleases and host defense: An evolving story" (https://www.
ncbi.nlm.nih.gov/pmc/articles/PMC2692241) . Journal of Leukocyte Biology. 83 (5): 1079–87.
doi:10.1189/jlb.1107725 (https://doi.org/10.1189%2Fjlb.1107725) . PMC 2692241 (https://www.n
cbi.nlm.nih.gov/pmc/articles/PMC2692241) . PMID 18211964 (https://pubmed.ncbi.nlm.nih.gov/1
8211964) .

External links

Two-phase wash t o solve t he ubiquit ous cont aminant -carryover problem in commercial
nucleic-acid ext ract ion kit s (ht t ps://www.nat ure.com/art icles/s41598-020-58586-3) ; by
Erik Jue, Daan Wit t ers & Rust em F. Ismagilov; Nat ure, Scient ific report s, 2020.
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