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 and section samples (with DNase)
Applications	RT-PCR, quantitative RT-PCR, Northern hybridization, Poly A purification, nucleic acid protection and in vitro translation
Purification method	Mini spin column
Purification technology	Silica technology, DNase
Process method	Manual (centrifugation or vacuum)
Sample type	FFPE tissue sample
Sample amount	6mg
Yield	20μg
Elution volume	≥10μl
Time per run	≤60 minutes
Liquid carrying volume per column	800µl
Binding yield of column	100µg

Principle

This product is based on silica column purification. Remove paraffin by Buffer DPS. Sample lysis with proteinase 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 was finally eluted with low-salt buffer.

Advantages

• 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
• Efficient DNA removal – unique method to effectively remove genomic DNA

Kit Contents

Contents	R414402	D414403
Purification Times	50 Preps	250 Preps
HiPure RNA Micro Columns	50	250
2ml Collection Tubes	50	250
Buffer DPS	60 ml	250 ml
Buffer FRL	15 ml	60 ml
Buffer RLC	15 ml	60 ml
Buffer RWC*	10 ml	50 ml
Buffer RW2*	20 ml	2 x 50 ml
DNase I	600 µl	5 x 600 µl
DNase Booster Buffer	1.5 ml	6 ml
Protease Dissolve Buffer	1.8 ml	10 ml
Proteinase K	24 mg	120 mg
RNase Free Water	10 ml	20 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 room temperature (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.

Introduction

This product is suitable for extracting RNA from anticoagulant blood, lymphocytes, buffy coat, bone marrow, cultured cells and other samples.

Details

Specifications

Features	Specifications
Main Functions	Isolation total RNA from 1-1.5ml whole blood, 0.5-1ml bone marrow, buffy coat
Applications	RT-PCR, cDNA synthesis, second generation sequencing
Purification method	Polydisperse magnetic beads
Purification technology	Magnetic beads technology
Process method	Manual or automatic
Sample type	anticoagulant blood, lymphocytes, buffy coat, bone marrow, cultured cells
Sample amount	1-1.5ml whole blood, 0.5-1ml bone marrow
Yield	1-30μg

Principle

This product is suitable for extracting RNA from anticoagulant blood, lymphocytes, buffy coat, bone marrow, cultured cells and other samples. This product is based on the purification method of high binding magnetic particles. The sample is lysed and digested by lysis buffer and protease, and RNA/DNA is released into the lysis buffer. Add binding solution and magnetic particles to adsorb RNA/DNA, while proteins are not adsorbed and removed. The particles adsorbed with DNA/RNA are washed with washing buffer to remove proteins and other impurities, then washed with ethanol to remove salt, and finally digested with DNase to remove DNA. RNA is recovered by adding binding solution, and finally the RNA is eluted with low salt buffer. The eluted RNA can be directly used for experiments such as RT-PCR,NGS and virus detection.

Advantages

• High purity – OD 260 / 280 :1.9-2.0, OD 260 / 230 :1.5-2.0
• Economy – less than 50% of the price of top brands
• Automatic – extraction without manual participation, saving time and effort
• High yield – nucleic acid recovery up to 95%
• General – samples include whole blood, serum, plasma, lymphocyte suspension, bone marrow, cultured cells, etc.

Kit Contents

Contents	R661101	R661102	R661103
Purification Times	48 Preps	96 Preps	480 preps
10 x RBC Lysis Buffer	50 ml	2 x 50 ml	4 x 100 ml
Proteinase K	12 mg	24 mg	120 mg
Protease Dissolve Buffer	1.8 ml	1.8 ml	10 ml
DNase I	600 μl	2 x 600 μl	10 x 600 µl
DNase Buffer	20 ml	30 ml	150 ml
MagPure Particles N	1.2 ml	2.5 ml	11 ml
Buffer RTL	30 ml	60 ml	300 ml
Buffer ALB2	40 ml	60 ml	300 ml
Buffer MW1*	22 ml	44 ml	220 ml
Buffer MW2*	20 ml	50 ml	2 x 100 ml
RNase Free Water	10 ml	20 ml	60 ml

Storage and Stability

DNase I should be shipped with ice pack or dry ice and stored at -20°C upon arrival. MagPure Particles N and Proteinase K should be stored at 2–8°C upon arrival. However, short-term storage(up to 8 weeks) at room temperature (15–25°C) does not affect their performance.The remaining kit components can be store at room temperature and are stable for up to 18 months under these conditions.

This product is suitable for extracting RNA from anticoagulant blood, lymphocytes, buffy coat, bone marrow, cultured cells and other samples.

The Flex caddy allows the Heater-Shaker GEN1 to be installed on the Opentrons Flex deck and sit below the deck, enabling below-deck cable routing and labware placed on top of modules to remain closer to the deck surface. The calibration adapter is used to calibrate the location of the Heater-Shaker. Caddies and calibration adapters are specific to the type of module.

Note: This item does not include the Heater-Shaker. The module is available with or without the Flex Caddy and Calibration tool: Heater-Shaker Module GEN1

The Flex caddy allows the Heater-Shaker GEN1 to be installed on the Opentrons Flex deck and sit below the deck, enabling below-deck cable routing and labware placed on top of modules to remain closer to the deck surface. The calibration adapter is used to calibrate the location of the Heater-Shaker. Caddies and calibration adapters are specific to the type of module.

Note: This item does not include the Heater-Shaker. The module is available with or without the Flex Caddy and Calibration tool: Heater-Shaker Module GEN1