Norgen’s CleanAll DNA/RNA Clean-Up and Concentration Micro Kit provides a rapid method for the purification, cleanup and concentration of RNA or DNA from different isolation methods or upstream applications. The kit can be used as an alternative to organic extraction and ethanol precipitation to clean up various enzymatic reactions.
The CleanAll Kit purifies RNA from phenol/guanidine-based protocols or from various upstream enzymatic reactions such as DNase treatment, labeling and in vitro transcription. Furthermore, endotoxins can be removed from previously purified RNA solutions.
The kit can be used to clean up DNA from digestions, ligations, PCR reactions, labeling reactions, DNA modification reactions and staining. The kit also provides a protocol for the rapid removal of endotoxins from previously purified DNA down to 0.1 EU/µg DNA or less.
Purify all sizes of RNA, from large mRNA and ribosomal RNA down to microRNA (miRNA) and small interfering RNA (siRNA).
Purify all sizes of DNA, from small PCR products, native or linearized plasmids, to genomic DNA. Using an alternative protocol, even oligonucleotides and smaller DNA fragments can be purified.
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Kit Specifications | |
Column Binding Capacity | 50 μg RNA 10 μg DNA |
Maximum Column Loading Volume | 600 μL |
Minimum Elution Volume | 20 μL |
Time to Complete 10 Purifications | 20 minutes |
Size of RNA Purified | All sizes, including small RNA (<200) |
Size of DNA Purified | ≥ 100 bp* |
Average Recovery | ≥90% for RNA ≥90% for DNA 100 bp to 10 kbp ≥75% for DNA ≥ 10 kbp |
* Applicable to smaller fragments or oligonucleotides with alternative protocol
Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years from the date of shipment.
Component | Cat. 23800 (50 preps) |
---|---|
Binding Buffer H | 60 mL |
Wash Solution K | 27 mL |
Elution Buffer L | 15 mL |
Micro Spin Columns | 50 |
Collection Tubes | 50 |
Elution Tubes (1.7 mL) | 50 |
Product Insert | 1 |
Sample type purification kit guide
The 16S V3-V4 Library Preparation Kit for Illumina consists of the reagents and components required for library preparation of the 16S V3-V4 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 V3-V4 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 V3-V4 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.
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 V3-V4 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. 70400 (24 preps) | Cat. 70410, 70420, 70430, 70440 (96 preps) |
---|---|---|---|
Amplicon PCR (PCR 1) | MGX Master Mix | 330 µL | 1,320 µL |
16S V3-V4 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 |
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