10x5ml Angle Rotor 

Overview

• Extract total RNA (including microRNA) from FFPE samples
• No phenol extraction step
• Includes DNase for optional on-column DNA removal
• Isolated RNA is of the highest quality and integrity
• Isolate a diversity of RNA species
• Purified RNA is suitable for a variety of downstream applications, including Small RNA Sequencing. Find out more information on Norgen’s NGS services
• Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix

Norgen’s FFPE RNA Purification Kits provide a rapid method for the isolation and purification of total RNA (including microRNA) from formalin-fixed paraffin-embedded (FFPE) tissue samples in as little as 1 hour. Using formalin to fix tissues leads to crosslinking of the RNA and proteins, and the process of embedding the tissue samples can also lead to fragmentation of the RNA over time. Norgen’s FFPE RNA Purification Kits provide conditions that allow for the partial reversing of the formalin modifications, resulting in a high quality and yield of RNA. These kits are able to purify all sizes of RNA, from large mRNA and ribosomal RNA down to microRNA (miRNA) and small interfering RNA (siRNA), depending on the age of the FFPE tissue as fragmentation of the RNA is known to occur over time. The RNA is preferentially purified from other cellular components without the use of phenol or chloroform.

FFPE RNA Purification Kit (Spin Column)

Maximum loading volume of 650 μL per column, and a maximum binding capacity of 50 μg per column.

FFPE RNA Purification 96-Well Kit (High Throughput)

Purification is based on 96-well column chromatography using Norgen’s proprietary resin as the separation matrix. Purification can be performed using either a vacuum manifold or centrifugation. Maximum loading volume of 400 μL per well, and a maximum binding capacity of 50 μg per well.

Details

Supporting Data

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Kit Specifications
Maximum Column Binding Capacity	Up to 50 µg RNA
Maximum Loading Volume Per Spin Column	650 µL
Size of RNA Purified	All sizes, including small RNA (< 200 nt)
Time to Complete 10 Purifications	1-4 hours*
Maximum Amount of Starting Material	5 slices of < 20 µm thick paraffin slices 25 mg of unsectioned block
Average Yield	Variable due to age of paraffin blocks ~2-3 µg of Total RNA per 1 mg of fresh FFPE hamster kidney

* Time required for purification varies by length of Proteinase K incubation and formalin crosslink-reversal

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Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. The DNAse I and Proteinase K should be stored at -20°C upon arrival. This kit is stable for 1 year from the date of shipment.

Component	Cat. 25300 (50 preps)	Cat. 25400 (2 x 96 preps)
Digestion Buffer A	25 mL	2 x 25 mL
Buffer RL	30 mL	2 x 30 mL
Enzyme Incubation Buffer	6 mL	2 x 6 mL
Wash Solution A	38 mL	2 x 38 mL
Elution Solution A	6 mL	2 x 20 mL
Proteinase K	12 mg	2 x 20 mg
DNase I	1 vial	2 x 500μL
Micro Spin Columns	50	–
96-Well Incubation Plate	–	2
96-Well Plate	–	2
Adhesive Tape	–	8
Collection Tubes	50	–
96-Well Collection Plate	–	2
Elution Tubes (1.7 mL)	50	–
96-Well Elution Plate	–	2
Product Insert	1	1

• Real time qPCR kit for AetA gene
• For screening aetokthonotoxin gene cluster
• Use in combination with Attogene Algae DNA isolation kit

Description

Not all cyanobacterial strains produce toxins. However, the toxin-producing strains cannot be distinguished from the nontoxin-producing strains by traditional light microscopy, commonlyused to monitor water bodies.  An alternative for the differentiation of potentially toxic strains from nontoxic strains is to use molecular methods to detect the presence of toxin biosynthetic genes. Such methods are already available and could be used for the detection and identification of potential microcystin and nodularin producers present in environmental samples (Attogene catalog number NA2024).

Screening for the toxin itself, can be very costly.  In turn, real time PCR for the detection of a gene region responsible for assembling  in cyanobacterial strains and environmental samples can be a key indicator for the prescense of cyanobacteria capable of expressing the aetokthonotoxin toxin.  Attogen has thus, designed primer pairs and probes targeting a the conserved gene region in order to enable the amplification and detection of several producer genera using real time PCR.  Screening for the toxin genes can save significant costs and act as a triage for samples needing to be analyzed for the toxin itself.

Cyanobacterial neurotoxin aetokthonotoxin (AETX), a peculiar pentabrominated biindole alkaloid implicated in fatal Vacuolar Myelinopathy.  This neurodegenerative disease was first recorded in 1994 during an outbreak of bald-eagle poisonings at De Gray Lake in Arkansas, USA.  AETX was experimentally confirmed to be produced by the true branching heterocytous cyanobacterium Aetokthonos hydrillicola.  The production of AETX is dependent on bromide (Br−) availability, and likely linked to its hyper-accumulation by the host plan.  Thus regular monitoring of A. hydrillicola (accompanied by assessment of Br− and AETX levels) is highly advisable to predict the possible threat of further VM outbreaks.

The cyanobacterial AetA gene which encodes the unique FAD-dependent halogenase involved in the pathway for AETX synthesis has been adapted to develop a -aetokthonotoxin specific quantitative PCR (qPCR) assay.

Real time qPCR kit for AetA gene
For screening aetokthonotoxin gene cluster
Use in combination with Attogene Algae DNA isolation kit

Character

High flatness sample shelf:

Silicone oil thermal conductive shelf, high-precision processing method of upper and lower clamping, shelf temperature uniformity can reach ±0.5°c

Integrated cavity:

Integrated chamber, the shelf and cold trap are located in the same chamber, the steam conduction is faster and the freeze-drying effect is better

Refrigerating and heating control system:

Using thermally conductive silicone oil with a high temperature range and precise PID control algorithm, the control accuracy can reach ±0.5°C. Sample shelf temperature range: -70°C~ +80°C

Freeze-drying endpoint testing system:

The pressure comparison method is used to determine the freeze-drying end point. The freeze-drying end point test can be automatically performed after the analytical drying stage to ensure that the moisture content of the material reaches the standard requirements.

Vacuum precise control and adjustment system:

The sublimation and analytical drying processes are controlled and adjusted with high precision to make the freeze-drying process warmer and the sample sublimation more reliable.

Pulse backfill system:

Three backfilling modes can be selected: slow, medium and fast, which avoids the problem of effective substances being blown away and lost due to the aeration of granular and flocculent materials after freeze-drying.

Manual automatic mode selection:

Manual mode is used to explore process parameters (strong human intervention), automatic mode is used in the mature stage of the process, one-click operation, simple and convenient

Data recording and analysis system:

Through the PC host computer, all operating parameters and background parameters of the equipment can be recorded in real time, and all parameter action changes and action changes of operating components can be recorded. 

Freeze-drying process recipe storage function:

The host can store 60 sets of fixed or user-defined freeze-drying process recipes, and the PC can store >10,000 programs (depending on the computer hard drive)

Eutectic point test function:

The eutectic point temperature of the product can be tested offline and online, and the spectrum can be analyzed manually or automatically (optional)

Remote control system:

The operating status of the device can be monitored remotely, and the device can be monitored in real time via WiFi or the cloud.

Mobile phone monitoring system:

You can monitor the operating status of the equipment with your mobile phone, which is simple and convenient (optional)

Cold trap defrost system:

The hot air defrosting method has high safety performance; the defrosting speed is fast, which solves the inconvenience caused by traditional immersion defrosting.

Parameter

Model	EX2	EX5	EX8	EX12
Freeze-area	0.24m²	0.38m²	0.96m²	1.45m²
Ice coagulating capacity	4kg	9kg	12kg	18kg
Shelf temperature	70℃~+80℃			-65℃~+70℃
Cold trap temperature	-70/-90℃	-70/-90℃	-70℃/-90℃	-70℃/-85℃
Cold trap volume	9L	15L	18L	30L
chamber type	single	double
Shelf size	275*400 mm	275*400 mm	365*465 mm	365*465 mm
Shelf spacing	100mm(adjustable)	100mm(adjustable	70mm(adjustable)	70mm(adjustable)
Sample quantity (4R Cillin bottle)	558	1395	2630	4230
Shelf heating and cooling method	Thermal conducting silicone oil medium(temperature resistance： -100℃~+600℃)
Chamber and shelf surface	High vacuum stainless steel tube/KF25-40
vacuum line	High vacuum stainless steel tube			KF50
External valve port	6/	12/	18/	一
equipment power	3.8kw	5.5kw	6.5kw	8kw
equipment size	700*588*1600 mm	1030*700*1630 mm	1030*700*1850 mm	1500*850*1900 mm
Capping mode	Hydraulic system gland device/no gland			hydraulic pressure
Suitable clean room	Suitable
Vacuum sensor type	Pirani Vacuum sensor/Capacitive vacuum sensor
Remote control and alarm	Support 2G/4G/GSM/Wi-Fi
Terminal judgment function	Pressure contrast method	Pressure test/pressure contrast method
Freeze dryer front door cover	Stainless steel/Plexiglass			Stainless steel
Inert gas filling	(Optional) Sufficient amount and time for the program to run itself
User function customization	Can be customized and modified according to user needs
sterilization method	H2O2/steam sterilization