We found a match
Your institution may have access to this item. Find your institution then sign in to continue.
- Title
Comparative assessment of four RNA extraction methods and modification to obtain high-quality RNA from Parthenium hysterophorus leaf.
- Authors
Ahmad, Javed; Baig, M.; Ali, Arlene; Al-Huqail, Asma; Ibrahim, M.; Qureshi, M.
- Abstract
Isolation of high-quality RNA from weed plants such as Parthenium hysterophorus is a difficult task due to the hindrance caused by numerous secondary metabolites. Such metabolites not only affect the quality and yield of RNA, but also limit the quality of downstream applications. Therefore, the present study was undertaken to design a protocol for yielding RNA with better quality and quantity from P. hysterophorus leaf which could be suitable for functional genomics. To achieve the objective, four different important RNA extraction protocols, viz. acid guanidinium thiocyanate-phenol-chloroform, phenol-LiCl precipitation, TRIzol, and PVP-ethanol were tested. The PVP-ethanol method proved to be best among the tested protocols. This method was further modified for obtaining improved quality and yield of RNA. The modified method successfully enhanced the yield of RNA from 280 to 334 µg g fresh weight. The absorbance ratio ( A / A ) was in the purity range of 1.9 that indicated the good quality of RNA. To prove the feasibility of the extracted RNA in PCR-based cDNA synthesis, actin transcripts were targeted and successfully amplified using suitable primers. The improved protocol thus not only improved the yield and quality of RNA, but also gave better results in reverse transcriptase PCR.
- Subjects
RNA analysis; PARTHENIUM hysterophorus; GUANIDINE; CHLOROFORM; ANESTHETICS
- Publication
3 Biotech, 2017, Vol 7, Issue 6, p1
- ISSN
2190-572X
- Publication type
Article
- DOI
10.1007/s13205-017-1003-3