GSM1435111
GSM1435111: mtr1; Medicago truncatula; miRNA-Seq
Trizol reagent was used for total RNA extraction and the low molecular weight RNA was isolated by using mirVana kit. Total RNA (1µg) was ligated to adenylated 3’ HD adapter by using T4 RNA ligase 2 (truncated T4 RNA ligase). The ligated product was cleaned and concentrated by Zymo RNA cleaning and concentrating kit. The remaining adapter was de-adenylated by adenylase and degraded by exonuclease. Then the 3’ ligated products were ligated to 5’ HD adapter by using T4 RNA ligase 1, and the reaction product was further cleaned and concentrated by the Zymo RNA cleaning and concentrating kit. The concentrated RNA being ligated to both 5’ and 3’ HD adapters were reverse transcribed into cDNA and the cDNA was amplified by using Phusion DNA polymerase. The amplified products were run on 6-8% of PAGE gel. The band around the size of 147bp was cut and the PCR products, the library, were extracted from the gel and were sent for sequencing.
GEO
GSM1435111
GEO Accession
GSM1435111