Rat kidney was stabilized in RNAlater RNA Stabilization Reagent, and 25 mg samples were disrupted in QIAzol Lysis Reagent for 2 x 5 minutes at 25 Hz using the TissueLyser II. Total RNA containing miRNA was purified using the miRNeasy Mini Kit, using either the manual procedure (Manual) or the automated procedure with the QIAcube (QIAcube). Real-time RT-PCR using the miScript System was carried out to detect 2 different miRNAs, miR-16 and miR-25.
Frozen plant leaves were disrupted using the TissueLyser II (2 x 1 minute). RNA was purified using the RNeasy Plant Mini Kit and analyzed on a 1.2 % formaldehyde agarose gel. The ribosomal RNA bands were sharp and of equal intensity, indicating reproducible purification of intact RNA. T: tomato (100 mg); A: arabidopsis (25 mg); C: cotton (100 mg); M: maize (100 mg); R: rape (100 mg).
Various rat tissues (25 mg each)
stabilized in Allprotect Tissue Reagent were disrupted using the TissueLyser LT or TissueLyser II.
Total protein was purified using the Qproteome Mammalian Protein Prep Kit and then
analyzed by western blotting. Transfer membrane stained with Ponceau S, and western
blot. LT: TissueLyser LT; II: TissueLyser II; M: markers.
tissues (25 mg each) stabilized in Allprotect Tissue Reagent
were disrupted using the TissueLyser LT or TissueLyser II.
DNA was purified on the QIAcube using the DNeasy Blood
& Tissue Kit, and then used in PCR with the HotStarTaq Plus
Master Mix Kit and a PGK1 primer system. The 120 bp PCR
product was analyzed on the QIAxcel using the QIAxcel
DNA High Resolution Kit. LT: TissueLyser LT; II: TissueLyser II;
plant tissues (100 mg each) were disrupted in precooled
adapters using the TissueLyser LT or TissueLyser II. DNA was
purified on on the QIAcube using the DNeasy Plant Mini Kit
and then analyzed on an agarose gel. LT: TissueLyser LT;
II: TissueLyser II; M: markers.