However, miRNAs in plasma are surprisingly stable (3) despite the presence of RNAases
0) and treated with 1 [micro]L of RNAase
A (10mg [mL.
2 unit RNAase
H was used to remove RNA hybridized with cDNA for 30 min at 37[degrees]C.
Each of the tubes received 3 [micro]l of the RNAase
solution from Wizard and these were mixed by inversion.
In the nucleus, miRNAs are transcribed as primary pri-miRNA transcripts and then are processed to form the precursor pre-miRNA stem loop structure before transportation into the cytoplasm [where they are cleaved by the Dicer RNAase
III endonuclease and produce mature miRNA (21-23 nucleotides] (20).
2 70% (v/v) ethanol Chloroform- isoamyl alcohol (24: 1 v/v) 10 mg/ml RNAase
TE: 10 mM Tris-HCl pH 8.
5 mM of each nucleotide triphosphate, 40 units of RNAase
inhibitor (Ambion, USA), 50-100 ng of Random Hexamers (Amersham Pharmacia, UK).
Importantly, for surface mucous RAPD analysis, the RNAase
treatment of nucleic acid was a necessary step, whereas the inclusion of proteinase K was not.
5S) small cytoplasmic required for protein RNA secretion; function is similar to 7S in eukaryotes RNA subunit of The enzyme RNase P is 350-410 nt RNAase
P necessary for processing of bacterial tRNAs to their functional length; the RNA molecule is the catalytic subunit and the protein subunit acts as a cofactor and helps tRNA binding snRNA Components of the There are five major small nuclear RNA eukaryotic splicosome, species of snRNA, termed which is necessary for U1, U2, U4, U5, and U6, splicing and removal approximately 100-600 nt of introns in the premRNA molecules snoRNA Components of the There are 77 genes small nucleolar nucleolus in encoding snoRNA in S.
The second strand was synthesized using second strand mix, which contained both RNAase
H and DNA polymerase.
The cDNA synthesis was carried out using AMVRTase (10 U) at 42[degrees]C for 90 min, in a total volume of 25[micro]l containing RNAase
inhibitor (25 U), 1 mM each dNTP and downstream primer (15 pmol).
Zn is an essential micronutrient for plant growth and under deficiency conditions, protein synthesis is dramatically reduced (due to the relationship with RNAase
activity); causes delay and reduction in plant growth; small and poorly shaped leaves; short internodes; formation of leaves in rosettes; internerval chlorosis (due to the involvement of Zn in chlorophyll formation), and necrosis at the apical root meristem (Broadley et al.
A 400-[micro]L plasma sample was first lysed using the Qiagen protease, which is free of DNAase and RNAase
activity, and with buffering conditions (salts and pH) that allow the binding of only DNA to the QIAamp membrane, excluding proteins and other contaminants that can inhibit PCR.
Pelleted cells were suspended in propidium iodide solution (500[micro]g/ml made in RNAase
A containing buffer) for 30 min.
DNA-free total RNA (4 Ag) was extracted from plants of control and Al-treated brassica was reverse-transcribed in a 20 [micro]l final reaction volume containing 1[micro]l oligo-dT, 1[micro]l random primer, 4 [micro]l reactant buffer, 1 [micro]l dNTP, 0/5 [micro]l RNAase
inhibitor, 1 [micro]l reverse transcriptase.