Isolation and Purification of Poly(A)+ and Poly(A)- RNAs from Heat-Treated Pear Leaves

Poly(A)⁺ and Poly(A)⁻ RNAs were isolated from the total RNAs of heat-treated pear leaves using a polyA Spin^TM mRNA Isolation Kit (New England Biolabs, Ipswich, MA, USA). T4 polynucleotide kinase (New England Biolabs), RNA 5′ pyrophosphohydrolase (New England Biolabs), and 5′–3′ exoribonuclease (New England Biolabs) were used for the RNA digestion, according to a previous study^{58 (link)}. After digestion, the RNAs were purified using the modified cetyltrimenthyl ammonium bromide (CTAB) method^{59 (link)} and subjected to RT-PCR. Primer sequences are provided in Supplementary Data 1.

Free full text: Click here

Zhang Y., Wang S., Li W., Wang S., Hao L., Xu C., Yu Y., Xiang L., Li T, & Jiang F. (2022). A long noncoding RNA HILinc1 enhances pear thermotolerance by stabilizing PbHILT1 transcripts through complementary base pairing. Communications Biology, 5, 1134.

Publication 2022

polyA SpinTM mRNA Isolation Kit T4 polynucleotide kinase RNA 5′ pyrophosphohydrolase

5 3 exoribonuclease Ammonium bromide Digestion Isolation Pear Poly a Poly a rnas Polya mrna Polynucleotide kinase Primer Rnas Rt pcr

Corresponding Organization :

Other organizations : China Agricultural University

Top 5 similar protocols

Variable analysis

independent variables

Heat treatment of pear leaves

dependent variables

Isolation of Poly(A)+ and Poly(A)- RNAs from total RNAs of heat-treated pear leaves

control variables

Use of polyA Spin™ mRNA Isolation Kit (New England Biolabs) for RNA isolation
Use of T4 polynucleotide kinase, RNA 5′ pyrophosphohydrolase, and 5′–3′ exoribonuclease (all from New England Biolabs) for RNA digestion
Use of modified cetyltrimenthyl ammonium bromide (CTAB) method for RNA purification
Subsequent RT-PCR analysis

positive controls

Not specified

negative controls

Not specified

Annotations

Based on most similar protocols

Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.

As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!