Isolation of high-quality RNA from Platycladus orientalis and other Cupressaceae plants
Platycladus orientalis has a lifespan of several thousand years in China, making it a good plant in which to study aging at the molecular level, but this requires sufficient quantities of high-quality P. orientalis RNA. However, no appropriate methods have been reported for total RNA isolation from...
Guardado en:
| Autores principales: | , , , , |
|---|---|
| Lenguaje: | English |
| Publicado: |
Pontificia Universidad Católica de Valparaíso
2016
|
| Materias: | |
| Acceso en línea: | http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582016000500004 |
| Etiquetas: |
Agregar Etiqueta
Sin Etiquetas, Sea el primero en etiquetar este registro!
|
| id |
oai:scielo:S0717-34582016000500004 |
|---|---|
| record_format |
dspace |
| spelling |
oai:scielo:S0717-345820160005000042016-10-26Isolation of high-quality RNA from Platycladus orientalis and other Cupressaceae plantsChang,ErmeiZhao,YuxinWei,QianchunShi,ShengqingJiang,Zeping Cupressaceae plants Polysaccharides Polyphenols Total RNA isolation Two-step removal of polyphenols and polysaccharides Platycladus orientalis has a lifespan of several thousand years in China, making it a good plant in which to study aging at the molecular level, but this requires sufficient quantities of high-quality P. orientalis RNA. However, no appropriate methods have been reported for total RNA isolation from P. orientalis leaves. The TRIzol method did not extract RNA, while cetyltrimethylammonium bromide, sodium dodecyl sulfate-phenol, and plant RNAout kit (Tianz, Inc., China) protocols resulted in low yields of poor quality RNA. Isolating total RNA using the Spectrum Plant Total RNA Kit (Sigma-Aldrich, St. Louis, MO, USA) resulted in a high-quality product but a low yield. However, the two-step removal of polyphenols and polysaccharides in the improved plant RNAout kit protocol resulted in the isolation of RNA with a 28S:18S rRNA ratio of band intensities that was ~2:1, the A260/A280 absorbance ratio was 2.03, and the total RNA yield from P. orientalis leaves was high. This protocol was tested on different P. orientalis tissues of different ages and on leaves of five other Cupressaceae plants. The total RNAs were successfully used in complementary DNA synthesis for transcriptome sequencing and would be suitable to use in additional experiments. The results of this study will benefit future studies in Cupressaceae plants.info:eu-repo/semantics/openAccessPontificia Universidad Católica de ValparaísoElectronic Journal of Biotechnology v.19 n.5 20162016-09-01text/htmlhttp://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582016000500004en10.1016/j.ejbt.2016.08.003 |
| institution |
Scielo Chile |
| collection |
Scielo Chile |
| language |
English |
| topic |
Cupressaceae plants Polysaccharides Polyphenols Total RNA isolation Two-step removal of polyphenols and polysaccharides |
| spellingShingle |
Cupressaceae plants Polysaccharides Polyphenols Total RNA isolation Two-step removal of polyphenols and polysaccharides Chang,Ermei Zhao,Yuxin Wei,Qianchun Shi,Shengqing Jiang,Zeping Isolation of high-quality RNA from Platycladus orientalis and other Cupressaceae plants |
| description |
Platycladus orientalis has a lifespan of several thousand years in China, making it a good plant in which to study aging at the molecular level, but this requires sufficient quantities of high-quality P. orientalis RNA. However, no appropriate methods have been reported for total RNA isolation from P. orientalis leaves. The TRIzol method did not extract RNA, while cetyltrimethylammonium bromide, sodium dodecyl sulfate-phenol, and plant RNAout kit (Tianz, Inc., China) protocols resulted in low yields of poor quality RNA. Isolating total RNA using the Spectrum Plant Total RNA Kit (Sigma-Aldrich, St. Louis, MO, USA) resulted in a high-quality product but a low yield. However, the two-step removal of polyphenols and polysaccharides in the improved plant RNAout kit protocol resulted in the isolation of RNA with a 28S:18S rRNA ratio of band intensities that was ~2:1, the A260/A280 absorbance ratio was 2.03, and the total RNA yield from P. orientalis leaves was high. This protocol was tested on different P. orientalis tissues of different ages and on leaves of five other Cupressaceae plants. The total RNAs were successfully used in complementary DNA synthesis for transcriptome sequencing and would be suitable to use in additional experiments. The results of this study will benefit future studies in Cupressaceae plants. |
| author |
Chang,Ermei Zhao,Yuxin Wei,Qianchun Shi,Shengqing Jiang,Zeping |
| author_facet |
Chang,Ermei Zhao,Yuxin Wei,Qianchun Shi,Shengqing Jiang,Zeping |
| author_sort |
Chang,Ermei |
| title |
Isolation of high-quality RNA from Platycladus orientalis and other Cupressaceae plants |
| title_short |
Isolation of high-quality RNA from Platycladus orientalis and other Cupressaceae plants |
| title_full |
Isolation of high-quality RNA from Platycladus orientalis and other Cupressaceae plants |
| title_fullStr |
Isolation of high-quality RNA from Platycladus orientalis and other Cupressaceae plants |
| title_full_unstemmed |
Isolation of high-quality RNA from Platycladus orientalis and other Cupressaceae plants |
| title_sort |
isolation of high-quality rna from platycladus orientalis and other cupressaceae plants |
| publisher |
Pontificia Universidad Católica de Valparaíso |
| publishDate |
2016 |
| url |
http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582016000500004 |
| work_keys_str_mv |
AT changermei isolationofhighqualityrnafromplatycladusorientalisandothercupressaceaeplants AT zhaoyuxin isolationofhighqualityrnafromplatycladusorientalisandothercupressaceaeplants AT weiqianchun isolationofhighqualityrnafromplatycladusorientalisandothercupressaceaeplants AT shishengqing isolationofhighqualityrnafromplatycladusorientalisandothercupressaceaeplants AT jiangzeping isolationofhighqualityrnafromplatycladusorientalisandothercupressaceaeplants |
| _version_ |
1718441934476804096 |