Sample type purification kit guide
The ITS1 Library Preparation Kit for Illumina consists of the reagents and components required for library preparation of the fungal target ITS1 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 XPMagneticBeads(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 ITS1 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 ITS1 region of the fungal DNA.The post-PCR reaction is then cleaned up using AMPure XPbeads. 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 XPbeads. The libraries are then ready for quantification, pooling and sequencing.
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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 ITS1 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 will remain stable for at least 1 year when stored at the specified storage conditions.
Propargyl-PEG8-t-butyl ester is a PEG reagent with an alkyne group and a t-butyl group. The alkyne group enables the formation of triazole linkage with azide-bearing compounds or biomolecules; copper is required as a catalyst. The t-butyl group can be hydrolyzed under acidic conditions. The hydrophicility of the molecule is improved by having 8 units of PEG. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
Propargyl-PEG8-t-butyl ester is a PEG reagent with an alkyne group and a t-butyl group. The alkyne group enables the formation of triazole linkage with azide-bearing compounds or biomolecules; copper is required as a catalyst. The t-butyl group can be hydrolyzed under acidic conditions. The hydrophicility of the molecule is improved by having 8 units of PEG. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
Norgen’s Plant/Fungi Total RNA Purification Kit provides a rapid method for the isolation and purification of total RNA, including virus and viroid RNA, from a wide range of plants. Total RNA can be purified from fresh or frozen plant tissues, plant cells or filamentous fungi samples using this kit. All sizes of RNA are purified, including microRNA (miRNA) . The procedure is rapid and convenient.
The RNA is purified without the use of phenol or chloroform. The purified RNA is of the highest quality, and can be used in a number of downstream applications including real time PCR, reverse transcription PCR, Northern blotting, RNase protection and primer extension, and expression array assays.
Norgen’s Plant/Fungi Total RNA Purification Kit is also available in a 96-well (High Throughput) format for high throughput applications. Purification with the 96-well plates can be performed using either a vacuum manifold or centrifugation.
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| Kit Specifications – Spin Column | |
| Maximum Column Binding Capacity | 50 μg |
| Maximum Column Loading Volume | 650 μL |
| Size of RNA Purified | All sizes, including small RNA (< 200 nt) |
| Maximum Amount of Starting Material: Plant Tissues Plant Cells Fungi | 50 mg 1 x 106 cells 50 mg (wet weight) |
| Average Yield* 50 mg Tomato Leaves 50 mg Tobacco Leaves 50 mg Plum Leaves 50 mg Grape Leaves 50 mg Peach Leaves | 60 μg 60 μg 32 μg 35 μg 30 μg |
| Time to Complete 10 Purifications | 30 minutes |
* Yield will vary depending on the type of sample processed.
* Yield will vary depending on the type of sample processed.
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.
Tobacco (Nicotiana tabacum)
Tomato (Lycopersicon esculentum)
Pepper (Capsicum annuum)
Potato (Solanum tuberosum)
Arabidopsis thaliana1
Peach (Prunus persica)
Apple (Malus sp.)
Pear (Pyrus sp.)
Grape vine (Vitis sp.)
Plum (Prunus sp.)
Palm (Arecaceae)
Pine needle (Pinaceae)
Strawberry
Raspberry
Blackberry
Herbs
Persimmon (Ebenaceae)
Potato tuber (Solanum)
Plum fruit
Citrus
Vanilla bean
Cotton (Gossypium)
Mangrove
Chrysanthemum
Grape berry skin
Kiwi leaves
Peach (fruits and flowers)
Soy bean (legume)
Eastern White Red Cedar
Corn leaves
Cucumber leaves
Aspergillus niger
Mucor racemosus
Cladosporium cladosporioides
Fusarium oxysporum
Penicillium sp.
Botrytis cinerea (Botryotinia fuckeliana)
Pichia sp.
Rhizopus oryzae
Alternaria tenuissima
| Component | Cat. 25800 (50 preps) | Cat. 31350 (100 preps) | Cat. 25850 (250 preps) | Cat. 31900 (192 preps) |
|---|---|---|---|---|
| Lysis Buffer C | 60 mL | 1 x 30 mL, 1 x 60 mL | 3 x 60 mL | 2 x 60 mL |
| Wash Solution A | 38 mL | 38 mL | 1 x 18 mL, 2 x 38 mL | 2 x 38 mL |
| Elution Solution A | 6 mL | 6 mL | 20 mL | 20 mL |
| Filter Columns | 50 | 100 | 250 | – |
| Spin Columns | 50 | 100 | 250 | – |
| 96-Well Plate | – | – | – | 2 |
| Adhesive Tape | – | – | – | 4 |
| Collection Tubes | 100 | 200 | 500 | – |
| 96-Well Collection Plate | – | – | – | 2 |
| Elution Tubes (1.7 mL) | 50 | 100 | 250 | – |
| 96-Well Elution Plate | – | – | – | 2 |
| Product Insert | 1 | 1 | 1 | 1 |