More Functional RNA Yield from FFPE Samples
The ability to extract RNA from FFPE samples allows correlation of disease state and tissue morphology with gene expression. Extraction of RNA from FFPE tissues is challenging because the formalin fixation process cross-links proteins and nucleic acids and introduces modifications that can inhibit subsequent analysis of RNA isolated from FFPE tissues.
The ReliaPrep™ FFPE Total RNA Miniprep System optimizes lysis conditions to reverse these RNA modifications without the need for overnight digestion. The system also uses a deparaffinization method that does not rely on xylene or other organic solvents. By eliminating the washing steps involved in xylene deparaffinization, this method can help retain small RNA fragments.
The protocol is complete in 1.5 hours with minimal hands-on time.
ReliaPrep™ FFPE Total RNA System Helps Preserve RNA Integrity
In this experiment, 5µl of purified RNA was used as template in a GoScript™ Reverse Transcription System reaction, and 5µl of the cDNA product was used as template for the multiplex PCR. Fragments from 80–290bp were detected.
Simple Protocol
Customer Data
Sample Type: Rat kidney, 2 years old. Four 10µm FFPE sections were purified together (total amount = 40µm) and diluted in 40µl. Eight microliters were used for gel electrophoresis.
Sample Type: Human liver tumor, fixed less than 1 year. One microliter out of 40µl total dilution of RNA was used for the gel.
Sample Type: Human kidney tumor, 2 months old. Two samples were extracted using the total RNA kit. Total dilution volume was 30µl, and the loading volume for agarose gel electrophoresis was 15µl. Purity: A260 / A280 ~2.0 A260 / A230 ~2.1 Concentration: 200ng/µl.
Protocols
Specifications
Catalog Number:
選択製品の構成品内容
| Item | Part # | Size | Available Separately |
|---|---|---|---|
|
Collection Tubes |
A130C | 1 × 10/pk | |
|
Mineral Oil |
A244A | 1 × 5ml | |
|
ReliaPrep™ FFPE Binding Columns |
A247A | 1 × 10/pk | |
|
Proteinase K (PK) Solution |
A505A | 1 × 250μl | View Product |
|
Nuclease-Free Water |
P119A | 1 × 1,250μl | View Product |
|
Lysis Buffer (LBA) |
Z101A | 1 × 1ml | |
|
DNase Buffer |
Z102A | 1 × 70μl | |
|
BL Buffer |
Z103A | 1 × 3.25ml | |
|
Wash Solution |
Z104A | 1 × 3ml | |
|
Elution Tubes |
Z200A | 2 × 5/pk | |
|
MnCl2, 0.09M |
Z318A | 1 × 250μl | |
|
DNase I (lyophilized) |
Z358A | 1 × 1 vial | View Product |
Blue Dye |
MC122A | 1 × 100μl |
選択製品の構成品内容
| Item | Part # | Size | Available Separately |
|---|---|---|---|
|
Collection Tubes |
A130A | 2 × 50/pk | |
|
Mineral Oil |
A244B | 1 × 50ml | |
|
ReliaPrep™ FFPE Binding Columns |
A247B | 2 × 50/pk | |
|
Proteinase K (PK) Solution |
A505B | 1 × 1.1ml | View Product |
|
Nuclease-Free Water |
P119E | 1 × 13ml | View Product |
|
Lysis Buffer (LBA) |
Z101B | 1 × 30ml | |
|
DNase Buffer |
Z102B | 1 × 700μl | |
|
BL Buffer |
Z103B | 1 × 32.5ml | |
|
Wash Solution |
Z104B | 1 × 30ml | |
|
Elution Tubes |
Z200C | 2 × 50/pk | |
|
MnCl2, 0.09M |
Z318B | 2 × 750μl | |
|
DNase I (lyophilized) |
Z358A | 4 × 1 vial | View Product |
Blue Dye |
MC122A | 1 × 100μl |
Resources
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