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Presto™ FFPE RNA Mini Kit (RF050, RF100) without DNase

RNA Extraction/Total RNA

The Presto™ FFPE RNA Mini Kit was designed specifically for purifying total RNA from formalin-fixed, paraffin-embedded (FFPE) tissue. Paraffin is initially dissolved with organic solvents such as Xylene followed by Proteinase K digestion. RNA in chaotropic salt is then bound by the glass fiber matrix of the spin column and DNA contamination is removed using an in column DNase treatment. Other contaminants are removed using Wash Buffer and the purified total RNA is eluted in RNase-free Water. High quality total RNA can be purified within 80 minutes without phenol extraction or alcohol precipitation. The purified RNA is ready for use in Real-time PCR, mRNA selection, cDNA synthesis and NGS.

DOCUMENTS

DESCRIPTION

Specifications (Cat. # RF050, RF100)

  • Sample: up to 25 mg sections of FFPE, 4~10 sections of 5~20 μm thick FFPE
  • High Yield: up to 50 μg of total RNA
  • Spin Columns: glass fiber membrane optimized for total RNA extraction
  • Individually packaged RNA spin columns and collection tubes, certified RNase and DNase-free
  • Storage: Storage: dry at room temperature (15-25°C)
  • Operation Time: within 80 minutes

APPLICATIONS

Real-time PCR, mRNA Selection, cDNA Synthesis, NGS

COMPONENTS

  • RT Buffer
  • RB Buffer
  • Proteinase K
  • W1 Buffer
  • Wash Buffer
  • RNase-free Water
  • RB Columns
  • 2 ml Collection Tubes

QUALITY CONTROL

The quality of the Presto™ FFPE RNA Mini Kit is tested on a lot-to-lot basis according to Geneaid's ISO-certified quality management system. Total RNA is isolated from 25 mg sections of FFPE, quantified with a spectrophotometer and analyzed by electrophoresis.

PrestoTM FFPE RNA Mini Kit Test Data

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Figure 1 .(A)Total RNA was isolated from 4 slices of chicken FFPE kidney section (5 μm thickness) using Geneaid PrestoTM FFPE RNA Mini Kit. The RNA integrity was analyzed by capillary electrophoresis (BIOptic, Qsep100). The RQN (RNA Quality Number) = 7.4 indicated the intact RNA was purified. However, due to effects of formalin fixation, embedding and storage, RNA from FFPE samples will usually show varying degrees of degradation. Typical RQN of RNA purified with PrestoTM FFPE RNA Mini Kit are in range of 2–6. (B)Since the RIN values from degraded FFPE samples are not a sensitive measure of RNA quality. Instead, the DV200 (percentage of RNA fragments that are greater than 200 nucleotides) developed by Illumina is used to assess FFPE RNA quality. In this case, the DV200 of purified RNA is 90.4%. The data showed that more than 90% of large RNA fragments in the purified RNA are excellent for successful library preparation. In general, the DV200 of purified RNA are in range of 30%-90% using Geneaid PrestoTM FFPE RNA Mini Kit.

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Figure 2 .High efficiency in real-time PCR.Total RNA was extracted from 2-5 slices of  5-20 μm thick FFPE sections from 15 normal endometrium patients using Geneaid PrestoTM FFPE RNA Mini Kit. The TET1 and GAPDH mRNA expression level were measured by real time one-step RT-PCR. The data showed that the purified RNA with excellent performance in downstream application such as RT-PCR.

Total RNA Extraction Kits Specifications

 
SpecificationsTotal RNA Mini Kit Blood Cultured CellTotal RNA Mini Kit TissueTotal RNA Mini Kit PlantPresto™ Mini RNA Bacteria KitPresto™ Mini RNA Yeast KitmiRNA Isolation KitGENEzol™ TriRNA Pure Kit
Cat. #RBD050
RBD100
RBD300
RTD050
RTD100
RTD300
RPD050
RPD100
RPD300
RBBD050
RBBD100
RBBD300
RBYD050
RBYD100
RBYD300
RMI050GZXD050
GZXD100
GZXD200
FormatSpin ColumnSpin ColumnSpin ColumnSpin ColumnSpin ColumnSpin ColumnTri Reagent Spin Column
Binding Capacity50 µg50 µg50 µg50 µg50 µg50 µg50 µg
Sampleblood, cellsanimal tissueplant tissuebacterial cellsyeast cellsanimal tissue, cellsblood,cells,
tissue, body fluids, bacteria
MicropestleN/AYesN/AN/AN/AYesN/A
Filter ColumnN/AYesYesN/AN/AN/AN/A
EnzymeDnase IDnase IDnase ILysozyme
Dnase I
Dnase IN/ADnase I
Typical Yield2-3 µg from 300 µl blood, 25-30 µg from 106 cells5-30 µg5-30 µg40-50 µg from 1x109 bacterial cells15-20 µg from 5x107 yeast cells5-30 µg5-30 µg
Elution Volume50 µl50 µl50 µl50 µl50 µl50 µl25-50 µl
Time<20 min.<15 min.<20 min.<20 min.<20 min.<30 min.<15 min.

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