Introduction

HiPure FFPE RNA Kit supplies a simple and rapid RNA extraction for Formalin-fixed, paraffin-embedded (FFPE) tissue and sections samples. This kit is based on silica gel column purification technology, no phenol-chloroform extraction or alcohol precipitation. The whole extraction only takes 30 minutes (not including digestion time). RNA can be directly used for downstream applications such as RT-PCR, Northern blot, vitro translation and other experiments.

Details

Specifications

Features	Specifications
Main Functions	Isolation total RNA from FFPE tissue
Applications	RT-PCR, cDNA synthesis, second generation sequencing
Products	RNA, miRNA
Purification method	Mini spin column
Purification technology	Silica technology, DNase
Process method	Manual (centrifugation or vacuum)
Sample type	FFPE slice, FFPE embedded tissue
Sample amount	Tissue: <6 mgFFPE: <6 slices
Yield	1-20μg
Elution volume	≥15μl
Time per run	≤30 minutes (after digestion)
Liquid carrying volume per column	800μl

Principle

This product is based on silica column purification. Remove paraffin by Buffer DPS. Sample lysis withproteinase K digestion requires only 15 minutes. After lysis, samples are incubated at 80ºC for 15 minutes. Transfer to an adsorption column and RNA is adsorbed on the membrane, while protein is not adsorbed and is removed with filtration. After washing proteins and other impurities, RNA wasfinally eluted with low-salt buffer.

Advantages

• Efficient removal of DNA – unique genomic DNA removal column without DNase treatment
• High quality – high purity total RNA can be directly used in various sensitive downstream applications
• Fast – several samples can be extracted in 60 minutes by column method
• Safe – no phenol chloroform extraction required
• Sensitive – RNA can be recovered at the level of PG

Kit Contents

Contents	IVD4144
Purification Times	50 Preps
HiPure RNA Micro Columns	50
2ml Collection Tubes	50
Proteinase K	24 mg
Protease Dissolve Buffer	1.8 ml
DNase I	600 μl
DNase Booster Buffer	1.5 ml
Buffer DPS	60 ml
Buffer FRL	15 ml
Buffer RLC	15 ml
Buffer RWC*	10 ml
Buffer RW2*	20 ml
RNase Free Water	10 ml

Storage and Stability

Proteinase K should be stored at 2–8°C upon arrival. DNase I should be stored at -20°C. However, short-term storage (DNase I up to 1 weeks, Proteinase K up to 8 weeks) at room temperature(15–25°C) does not affect their performance. The remaining kit components can be stored at roomtemperature (15–25°C) and are stable for at least 18 months under these conditions.

Experiment Data

HiPure FFPE RNA Kit supplies a simple and rapid RNA extraction for Formalin-fixed, paraffin-embedded (FFPE) tissue and sections samples. This kit is based on silica gel column purification technology, no phenol-chloroform extraction or alcohol precipitation. The whole extraction only takes 30 minutes (not including digestion time). RNA can be directly used for downstream applications such as RT-PCR, Northern blot, vitro translation and other experiments.

The Opentrons Flex Waste Chute GEN1 occupies position D3 and is used to accept waste such as tips, tip racks, and plates in high volumes. The gripper will pick up the consumable and dispose it into the chute based on your workflow needs. The waste chute is neccesary for all high-throughput workflows and can be paired with the Opentrons Flex Deck Expansion Set to maximize the throughput capabilities of the Opentrons Flex. The following consumables can disposed of into the waste chute:

– NEST 2 mL Deep Well Plate, V Bottom

– Opentrons Tough 0.2 mL 96-Well PCR Plate, Full Skirt

– NEST 96 well Plate Flat

– All Opentrons Flex tip racks

The Opentrons Flex Waste Chute GEN1 occupies position D3 and is used to accept waste such as tips, tip racks, and plates in high volumes. The gripper will pick up the consumable and dispose it into the chute based on your workflow needs. The waste chute is neccesary for all high-throughput workflows and can be paired with the Opentrons Flex Deck Expansion Set to maximize the throughput capabilities of the Opentrons Flex. The following consumables can disposed of into the waste chute:

– NEST 2 mL Deep Well Plate, V Bottom

– Opentrons Tough 0.2 mL 96-Well PCR Plate, Full Skirt

– NEST 96 well Plate Flat

– All Opentrons Flex tip racks

Description 

The PM5100 3-color Pre-Stained Protein Ladder High Range is a ready-to-use three-color protein standard with 14 pre-stained proteins covering a wide range of molecular weights from 10 to 245 kDa in Tris-Glycine Buffer (9 to 235 kDa in Bis-Tris (MOPS) buffer and 10 to 235 kDa in Bis-Tris (MES) buffer). Proteins are covalently coupled with different chromophores for easy identification of bands, with three reference proteins carrying enhanced intensity corresponding to a blue band at 20 kDa, green at 40 kDa, and red at 75 kDa, respectively, as separated on SDS-PAGE (Tris-Glycine buffer). The PM5100 3-color Pre-Stained Protein Ladder High Range is designed for monitoring protein separation during SDS-polyacrylamide gel electrophoresis, verification of Western transfer efficiency on membranes (PVDF, nylon, or nitrocellulose) and for approximating the size of proteins. 

Features

• Ready-to-use — Premixed with a loading buffer for direct loading, no need to boil.
• Three reference bands — 75 kDa (red), 40 kDa (green), and 20 kDa (blue)

Contents

Approximately 0.1~0.4 mg/ml of each protein in the buffer (20 mM Tris-phosphate (pH 7.5 at 25°C), 2% SDS, 0.2 mM DTT, 3.6 M urea, and 15% (v/v) glycerol). 

Quality Control 

Under suggested conditions, PM5100 ExcelBand™ 3-color Pre-Stained Protein Ladder High Range resolves 14 major bands in SDS-PAGE (Tris-Glycine, MOPS, and MES buffer) and after Western blotting to nitrocellulose membrane. 

Storage

4°C for 3 months
-20°C for long term storage

The PM5100 3-color Pre-Stained Protein Ladder High Range is a ready-to-use three-color protein standard with 14 pre-stained proteins covering a wide range of molecular weights from 10 to 245 kDa in Tris-Glycine Buffer (9 to 235 kDa in Bis-Tris (MOPS) buffer and 10 to 235 kDa in Bis-Tris (MES) buffer). Proteins are covalently coupled with different chromophores for easy identification of bands, with three reference proteins carrying enhanced intensity corresponding to a blue band at 20 kDa, green at 40 kDa, and red at 75 kDa, respectively, as separated on SDS-PAGE (Tris-Glycine buffer). The PM5100 3-color Pre-Stained Protein Ladder High Range is designed for monitoring protein separation during SDS-polyacrylamide gel electrophoresis, verification of Western transfer efficiency on membranes (PVDF, nylon, or nitrocellulose) and for approximating the size of proteins.