CO00020 Midstream Cassette 136x16mm, pink w/lens

The Opentrons Flex Gripper GEN1 attaches to the gantry and is capable of moving labware across the deck and on or off modules such as the Magnetic Block or Thermocycler. It can also access the additional slots to the side of the main deck to replenish tips or labware. The gripper can be used with any pipette configuration.

Included with the gripper is a calibration pin that enables automatic calibration to ensure positional accuracy. Currently, the gripper is optimized for use with the following labware items:

– NEST 2 mL 96 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 Gripper GEN1 attaches to the gantry and is capable of moving labware across the deck and on or off modules such as the Magnetic Block or Thermocycler. It can also access the additional slots to the side of the main deck to replenish tips or labware. The gripper can be used with any pipette configuration.

Included with the gripper is a calibration pin that enables automatic calibration to ensure positional accuracy. Currently, the gripper is optimized for use with the following labware items:

– NEST 2 mL 96 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

Overview

• Protocol optimized for DNA isolated from a diversity of samples including stool, soil, water, saliva, plant, urine, skin, and more
• Simple and quick workflow: library could be prepared in less than 5 hours
• Component of Norgen’s metagenomics workflow
• A single NGS run can be prepared with up to 384 unique dual-index libraries

Sample type purification kit guide 

The 16S V1-V2 Library Preparation Kit for Illumina consists of the reagents and components required for library preparation of the 16S V1-V2 amplicon libraries to be used for next-generation sequencing on Illumina platforms. All molecular reagents including primers, enzyme mixes, indexes, and buffers are provided. Instructions for PCR clean up with the AMPure XP Magnetic Beads (supplied by customer) are also included for rapid purification of nucleic acid products generated at two steps of the workflow. The library prep workflow could be used for purified DNA inputs from different sources including stool, soil, water, saliva, plant, urine, skin swab, vaginal swab, cheek swab, nasal swab, plasma/serum, tongue swab, gum swab, and others.

The 16S V1-V2 Library Preparation Kit for Illumina has a streamlined procedure that reduces the handling time such that the library prep procedure can be completed in approximately 4 hours (see diagram below). Input DNA is first subjected to targeted PCR to amplify the V1-V2 region of the DNA encoding 16S rRNA. The post-PCR reaction is then cleaned up using AMPure XP beads. Dual index primers are then added using a limited-cycle PCR. The indexed amplicons flanked by 5′ and 3′ barcoded adaptors are then cleaned using AMPure XP beads. The libraries are then ready for quantification, pooling and sequencing.

Details

Supporting Data

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Minimum amount of starting material:	2.5 µL of DNA (5 ng/µL)
Time to complete library preparation:	4 hours

Storage Conditions and Product Stability
Norgen’s 16S V1-V2 Library Prep Kit for Illumina is shipped as one kit box (for the 24 prep kit) or two sub-component kits (for the 96 prep kit). All kits should be stored at -20°C upon arrival.

All kit components should remain stable for at least 1 year when stored at the specified storage conditions.

Step	Component	Cat. 70100 (24 preps)	Cat. 70110, 70120, 70130, 70140 (96 preps)
Amplicon PCR (PCR 1)	MGX Master Mix	330 µL	1,320 µL
	16S V1-V2 Primer Mix	70 µL	280 µL
Index PCR (PCR 2)	Indexing Master Mix	660 µL	2 x 1,320 µL
	N7xx Index Primer	50 µL	50 µL
	S5xx Index Primer	70 µL	70 µL
PCR Clean-Up	Resuspension Buffer	2 x 1,250 µL	2 x 5,000 µL
	Nuclease-free water	1,250 µL	1 x 6,000 µL

Overview

• Isolate all sizes of circulating and exosomal RNA, including microRNA
• Versatile plasma/serum input ranges
• No phenol extractions
• No carrier RNA
• Bind and elute all RNA irrespective of size or GC content, without bias
• Concentrate circulating RNA and exosomal RNA into a flexible elution volume
• High quality, purified RNA is suitable for a variety of downstream applications, including Small RNA Sequencing. Find out more information on Norgen’s NGS services
• Compatible with Streck Cell-Free RNA BCT® Tubes
• Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix

These kits provide a fast, reliable and convenient method to purify and concentrate high quality, high purity and inhibitor-free cell-free circulating and exosomal RNA using a convenient spin column method. These kits can purify RNA from fresh or frozen serum or plasma samples prepared from blood collected on either EDTA or Citrate. Plasma samples prepared from blood collected on heparin should not be used, as heparin can significantly interfere with many downstream applications such as RT-PCR. The purified plasma/serum RNA is fully compatible with all downstream applications including PCR, qPCR, methylation-sensitive reverse transcription qPCR, reverse transcription PCR, Northern blotting, RNase protection and primer extension, expression array assays, and NGS.

Background

Plasma/Serum cell-free circulating RNA or exosomal RNA has the potential to provide biomarkers for certain cancers and disease states. Exosomes are 40 – 150 nm membrane vesicles, which are secreted by most cell types. Exosomes can be found in saliva, blood, urine, amniotic fluid and malignant ascitic fluids, among other biological fluids. Evidence has been accumulating recently that these vesicles act as cellular messengers, conveying information to distant cells and tissues within the body. These exosomes may play a functional role in mediating adaptive immune responses to infectious agents and tumours, tissue repair, neural communication and transfer of pathogenic proteins. For this reason exosomal RNAs may serve as biomarkers for various diseases including cancer. As the RNA molecules encapsulated within exosomes are protected from degradation by RNAses they can be efficiently recovered from biological fluids, such as plasma or serum.

Plasma/Serum RNA Purification Mini Kit

This kit can purify RNA from fresh or frozen serum or plasma samples prepared from blood collected on either EDTA or Citrate, from volumes ranging from 50 µL to 200 µL. The purified plasma/serum RNA is eluted in a flexible final volume of 10 µL to 25 µL.

Plasma/Serum RNA Purification Midi Kit

This utilizes a two column method, and can purify RNA from fresh or frozen serum or plasma samples prepared from blood collected on either EDTA or Citrate, from volumes ranging from 250 µL to 1.5 mL. The first column will handle the large volume input of bodily fluids that is followed by a concentration on a mini column for a final elution of 50 µL to 100 µL.

Plasma/Serum RNA Purification Maxi Kit

This kit can purify RNA from fresh or frozen serum or plasma samples prepared from blood collected on either EDTA or Citrate, from volumes ranging from 2 mL to 5 mL. The first column will handle the large volume input of bodily fluids that is followed by a concentration on a mini column for a final elution of 50 µL to 100 µL.

Isolate RNA after Purifying EVs and Exosomes

For Ultracentrifugation, Exoquick, and Filtration

Cat. #	Name	Elution Volume	Plasma/Serum	Urine	Cell-Culture Media
55000	Plasma/Serum RNA Purification Mini Kit	10 – 25 µL	50 µL – 1 mL	250 µL – 1 mL	5 – 10 mL
35300	Total RNA Purification Micro Kit	20 – 50 µL	1 – 4 mL	2 – 10 mL	10 – 20 mL
17200	Total RNA Purification Kit	50 – 100 µL	4 – 10 mL	11 – 30 mL	20 – 35 mL

Details

Supporting Data

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Kit Specifications
Sample Volume Range	50 to 200 μL
Anti-coagulant (for Plasma)*	EDTA or Citrate
Size of RNA Purified	All sizes, including small RNA (< 200 nt)
Minimum Elution Volume	10 μL
Maximum Elution Volume	25 μL
Time to Complete 10 Purifications	15-20 minutes
Average Yield**	Variable depending on specimen

*This kit is suitable for the isolation of RNA from fresh or frozen serum or plasma prepared from blood collected on either EDTA or Citrate. Plasma samples prepared from blood collected on heparin should not be used as heparin can significantly interfere with many downstream applications such as RT-PCR.

**Please check page 7 for Average Plasma/Serum Yields and Common RNA Quantification Methods.

Storage Conditions and Product Stability

All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment. It is recommended to warm Lysis Buffer A for 20 minutes at 60°C if any salt precipitation is observed.

Kit Components	Cat. 55000 (50 preps)	Cat. 56100 (20 preps)	Cat. 56200 (10 preps)
Lysis Buffer A	2 x 20 mL	100 mL	1 x 130 mL 1 x 30 mL
Wash Solution A	18 mL	1 x 38 mL 1 x 18 mL	38 mL
Elution Solution A	6 mL	6 mL	6 mL
Elution Buffer F	–	15 mL	15 mL
Micro Spin Columns	50	–	–
Mini Spin Columns	–	20	10
Midi Spin Columns	–	20	–
Maxi Spin Columns	–	–	10
Collection Tubes	50	20	10
Elution Tubes (1.7 mL)	50	20	10
Product Insert	1	1	1