A rapid and cost effective method in purifying small RNA

Abstract

Purification of RNA fragments from a complex mixture is a very common technique, and requires consideration of the time, cost, purity and yield of the purified RNA fragments. This study describes the fastest method of purifying small RNA with the lowest cost possible, without compromizing the yield and purity. The technique describes the purification of small RNA from polyacrylamide gel, resulting in a good yield of small RNA with minimum experimental steps in avoiding degradation of the RNA, obviating the use of ethidium bromide and phenol-chloroform extraction, as well as siliconized glass wools to remove the polyacrylamide gel particles. The purified small RNA is suitable for a wide variety of applications such as ligation, end labelling with radio isotope, RT-PCR (Reverse Transcriptase-PCR), Northern blotting, experimental RNomics study and also Systematic Evolution of Ligands by Exponential Enrichment (SELEX).

A high-performance waveguide-mode biosensor for detection of factor IX using PEG-based blocking agents to suppress non-specific binding and improve sensitivity

“…The RNA band was visualized under UV shadow and extracted from the excised gel-piece by using the crush-and-soak method. 28 The RNAs were precipitated in ethanol, dried under vacuum, and re-dissolved in RNase-free water, and the concentration was measured spectrophotometrically at 260 nm. Using the following DNA template, 5 0 -AGTAATACGACTCACTATAGGG TACCCCTGATATGGCGCATTACGAC-3 0 (T7 promoter region highlighted in italics), the complementary FIX aptamer was synthesized as described above using the same T7 forward primer and the reverse primer 5 0 -(T) 24 TACCCCTCCGTCG TAATGCGCCATAT-3 0 , for the negative reaction.…”

Reverse Electrochemical Sensing of FLT3-ITD Mutations in Acute Myeloid Leukemia Using Gold Sputtered ZnO-Nanorod Configured DNA Biosensors

“…Subsequent to ethanol precipitation of the PCR DNA amplicons, the templates were heat denatured at 95 • C with 8 M urea, resulting in complete separation of the complementary helical strands, being incapable of re-association. The denatured templates were PAGE purified (Figure 2C) and extracted using the crush-and-soak method [33].…”

Identification and characterization of miRNAs and target genes in developing flax seeds by multigroup analysis

“…The first strand of cDNA was synthesized by reverse transcription, and the double-strand library was obtained by polymerase chain reaction (PCR) amplification using cDNA as a template. In the small RNA library, the target fragments were purified by polyacrylamide gel [27]. qubit 2.0 was used to detect the library concentration.…”