I am extracting RNAs using Trizol combined with Purelink mini kit and I am doing post isolation DNAse treatment for making RNA free from genomic DNA. After DNAse treatment I do inactivate DNAse using EDTA and incubating at 70 degree Celsius for 10 minutes. Do I have have to do additional clean up again to before library preparation for RNAsequencing or I can proceed to library preparation after heat inactivation of DNAseI?
I have done this extra cleanup process using Qiagen RNAeasy minelute cleanup kit but it drastically reduced my RNA concentration. Please help me out.
@Sijan Pandit
You may use "Direct-zol RNA Miniprep Plus kit" from Zymo Reseaarch. It yields ultra-pure RNA without phenol carryover and RNA is ready for post NGS procedures.
You do not need separate DNase treatment as it included DNase I enzyme step during the spin column RNA extraction. You may get pure RNA with no DNA contamination.
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