To construct small RNA libraries, total RNA was extracted from leaves using Trizol reagent (Invitrogen, USA) according to the manufacturer's protocol as modified by LC Sciences (Houston, USA). The small RNA fraction (10-40 nt) was isolated by polyacrylamide gel electrophoresis and ligated with proprietary adaptors (Illumina). Short RNAs were then reverse-transcribed into cDNA by RT-PCR. Small RNA libraries were constructed for each maize line and sequenced using a Genome Analyzer GA-IIX (Illumina) following the manufacturer's instructions. Small RNA sequencing data was processed with the in-house program ACGT101-miR (LC Sciences, USA). Program parameters were set as previously described 36 (link) with modifications to adapt the program for plant miRNA prediction.
Protocol detail
Small RNA Library Construction
Partial Protocol Preview
This section provides a glimpse into the protocol.
The remaining content is hidden due to licensing restrictions, but the full text is available at the following link:
Access Free Full Text.
Corresponding Organization :
Other organizations : Anhui Agricultural University
Top 5 similar protocols
Variable analysis
independent variables
- Maize line
- Small RNA library construction
- Small RNA sequencing
- Total RNA extraction using Trizol reagent
- Small RNA fraction (10-40 nt) isolation by polyacrylamide gel electrophoresis
- Ligation of proprietary adaptors (Illumina)
- Reverse transcription of short RNAs into cDNA by RT-PCR
- Small RNA library construction for each maize line
- Small RNA sequencing using a Genome Analyzer GA-IIX (Illumina)
- Small RNA sequencing data processing with the in-house program ACGT101-miR (LC Sciences, USA)
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.
Sign up for free or login to display all annotations
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?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!