Storage term: stored at ≤-20℃,keep away from light, avoid heavy weight and repeated freezing and thawing.
Term of Validity: 14 months
Isothermal nucleic acid Principle Summary
The kit is based on room and constant temperature nucleic acid rapid amplification technology, its principle is that at room and constant temperature, the recombinase and primer form a protein/single-stranded nucleotide complex Rec/ssDNA, and invade the double-stranded DNA template with the help of auxiliary proteins and single-stranded binding protein SSB; then form a D-loop region at the invasion point and start to scan the DNA duplex, after finding the target region complementary to the primer and disintegration of the complex Rec/ssDNA, the polymerase also binds to the 3′ end of the primer to start the chain extension. The kit relies on the role of NFO enzyme and adds the designed specific molecular probes according to the template, and get the result by colloidal gold technology (sandwich method).
Technical Parameters:
Parameters
Details
Product Name
DNA Isothermal Amplification Kit NFO
Manufacturer
Amp-future
Storage Temperature
-20°C
Kit Components
Enzymes, Buffers ,Reagents
Packaging
48 Tests/box
Detection Limit
500-1000copies/µL
Shipping
ICE
Test Time
5-20mins
Isothermal nucleic acid Product Features
1/ High sensitivity and specificity, short reaction time.
2/ The reagent form is freeze-dried, stable and easy to operate.
3/ The reaction can be operated by metal bath and water bath pot without purchasing expensive PCR apparatus.
Isothermal nucleic acid Applications
Suitable for DNA isothermal rapid amplification kit(NFO type)
Primer: Require pair of nucleotide primers with the length of 25-35 bp.
DNA basic kit reaction temperature is 39 to 42℃ and time is 5-20 minutes.
Notes
1/ Please avoid nucleic acid contamination and set blank control during reaction due to the high sensitivity of the kit.
2/ Please take out the required quantity of MIRA reaction units for the experiment, and put the rest under storage conditions when performing the experiment.
Other Products
Small RNA Library Prep Kit for Illumina
Product Info
Document
Product Info
Overview
Optimized for low input RNA, especially from bodily fluids such as plasma or serum at 1ng of RNA
Simple and quick workflow: library could be prepared in less than 5 hours
No gel purification for selected types of samples
Protocol optimized for RNA isolated from different types of input, including liquid biopsies (blood, plasma, serum and urine)
Complements Norgen’s Best-in-Class Total RNA (including microRNA) Purification Technology
The Small RNA Library Prep Kit for Illumina consists of all the reagents and components required to generate small RNA libraries to be used for next-generation sequencing on an Illumina platform. All molecular reagents including adaptors, primers, enzyme mixes and buffers are provided. A purification module is also provided for rapid purification of nucleic acid products generated at various steps of the workflow. The purification module utilizes Norgen’s patent resin technology which enhances recovery of desired library intermediates or final products. The library prep workflow could be used for different forms of input including purified total RNA or enriched small RNA, as well as RNA from low content inputs such as plasma, serum and urine.
Storage Conditions and Product Stability Some components require storage at -20°C, 4°C or room temperature. See individual components and box labels for storage conditions.
Step
Component
Cat. 64600 (24 preps)
3′ AdaptorLigation to Template RNA
3′ Adaptor
30 µL
3′ Adaptor Ligation Master Mix
320 µL
T4 RNA Ligase 2 (Truncated)
35 µL
5′ Adaptor Ligation
5′ Adaptor
30 µL
5′ Adaptor Ligation Master Mix
320 µL
T4 RNA Ligase 1
35 µL
cDNA Synthesis from Ligated RNA Product
Reverse Primer
30 µL
cDNA Synthesis Master Mix
220 µL
TruScript ReverseTranscriptase
35 µL
PCR Amplification
2x NGS PCR Master Mix
1.32 µL
PCR Reverse Primer
81 µL
Forward Index Primer
Included in Small RNA Library Prep Forward Index Primers (# 64640 or # 64610)
Methylation Specific Bisulfite Seq Library Prep Kit
Product Info
Document
Product Info
Bisulfite seq is a well know technology to detect DNA methylation and several technologies such as WGBS, RRBS, MeDIP-Seq, and MSBS are used for whole genome DNA methylation analysis. DNA methylation is important for regulation of cell development, differentiation and gene expression in molecular biology, genetics and epigenetics. Most methylated cytosines are found at CpG sites, and 70-80% of cytosines are methylated. The number of CpG sites in human genome is around 28 million, which is less than 1% of the genome compared with 4.4% expected.
Whole genome bisulfite sequencing (WGBS) is the most effective method of DNA methylation analysis. The only limitation is the sequencing cost is very high because the whole genome is sequenced including all the non-methylated regions.
Reduced Representation Bisulfite Sequencing (RRBS) is the reduced representation of a smaller fraction of the methylated CpG sites. RRBS combines restriction enzyme digestion and bisulfite sequencing, and enriches the sequencing for methylated CpG sites. It is an efficient technology for estimate the whole genome methylation patterns at the single base level. Although this allows a higher coverage depth and reduces the sequencing cost, the limitation is only 10% of the methylated CpG sites are covered.
Methylated DNA Immunoprecipitation Sequencing (MeDIP-Seq) is another whole genome enrichment technique used for selection of methylated DNA. Using antibodies against 5-methylcytosine, methylated DNA is enriched from whole genomic DNA via immunoprecipitation. 5-methylcytosine antibodies are incubated with fragmented genomic DNA and precipitated, followed by DNA purification and sequencing. There are several drawbacks of MeDIP-Seq: 1. Low resolution (150~200 bp) as opposed to the single base resolution; 2. Non-specific interaction due to antibody specificity and selectivity. 3. Bias towards hypermethylated regions.
The Methylation Specific Bisulfite Seq (MSBS) Library Prep Kit (illumina platform) was developed for construction of NGS libraries for methylated CpG sites using bisulfite treated DNA (20 ng – 500 ng) as input. The kit enriches methylated CpG regions, thus significantly reduce the sequencing cost. The kit estimates the whole genome methylation patterns at the single base level since it is based on a bisulfite-seq technology.
It is known that bisulfite treatment of completed NGS libraries causes tremendous damage to the libraries. By using bisulfite treated DNA as input, the kit overcomes the significant library loss due to the bisulfite conversion. The kit contains a mixture of PCR polymerases that have high-fidelity amplification and uracil tolerance which is ideal for bisulfite treated DNA.
Methylation Specific Bisulfite Seq Library Prep Kit Workflow
Three index types are available for the kit:
Non-index (Cat.# 30101): Libraries do not have index.
Index (Cat.# 30102): Each primer contains a unique barcode sequence of 6 bases to identify the individual library. Library multiplexing capacity is up to 48 samples. Index information can be downloaded here.
Unique dual index (Cat.# 30103): The multiplexing of bisulfite sequencing library is up to 96 samples with unique dual indexes. We used a Four-Base Difference Index System to generate indexes that have at least 4 bases different from each other in the 8-base index. The index primers remove NGS errors including index cross-contamination, index hopping, reads mis-assignment etc. Index information can be downloaded here.
Methylation Specific Bisulfite Seq advantages
Enrichment of methylated CpG sites
Single-base resolution
Low cost for sequencing
Fast
Total time: 1.5 hours
Hands-on time: 10 minutes
Simple workflow
Bisulfite treated DNA as input: From 20 ng to 500 ng
MSBS Library Prep Kit enriches CpG sites
High methylation regions and low methylation regions in human genome.
High methylation region in human genome.
Low methylation region in human genome.
Sequencing setting: Single-end 35 cycles (Read 1, 35 bases) recommended To maximize the methylated CpG enrichment, we recommend to sequence the MSBS libraries with single end 35 cycles (read1, 35 bases). This is because the enriched methylated CpG sites are mainly located around the beginning of read 1 sequences. Shorter single end reads tend to have better methylated CpG enrichment.
Document
Bisulfite seq is a well know technology to detect DNA methylation and several technologies such as WGBS, RRBS, MeDIP-Seq, and MSBS are used for whole genome DNA methylation analysis. DNA methylation is important for regulation of cell development, differentiation and gene expression in molecular biology, genetics and epigenetics. Most methylated cytosines are found at CpG sites, and 70-80% of cytosines are methylated. The number of CpG sites in human genome is around 28 million, which is less than 1% of the genome compared with 4.4% expected.
Produced by reverse osmosis, ion-exchange, electro-deionization, ultrafiltration, UV treatment (UVC & 185 nm), 0.1 μm membrane filtration
Nuclease-free, protease-free, endotoxin-free
Suitable for molecular biology applications (qPCR, NGS etc.)
Suitable for cell culture applications
Produced in an ISO certified GMP laboratory
Water system is routinely monitored for compliance with QC standards
The Norgen water purification system has been designed in conjunction with experts from Millipore Sigma. The Ultrapure NFW is purified in a two-stage process. The first stage, to produce type II water, includes pre-treatment with activated charcoal and polyphosphate, this water then undergoes reverse osmosis, electro-deionization and ion exchange to remove contaminants as well as UV treatment for disinfection. The Pure water then flows to a second system in a clean room with an additional 4 layers of purification. After passing through an additional ion exchange column, the water is then subjected to Ultrafiltration to remove particulates, bacteria, & ionic contaminants down to trace level, the water is then treated with UV light (182 nm wavelength) to break down organic matter. Lastly a second Ultrafilter for the production of pyrogen-, nuclease- and bacteria-free water at 18.2 MΩ. This water is of utmost purity and suitable for direct use in molecular biology applications such as qPCR and NGS, as well as cell culture and other applications.
Product Specifications
pH
n/a (highly pure water does not contain enough ions for an exact pH determination.)
Recommended Storage
Room Temperature
Grade
Ultrapure (ASTM Type I)
Biological Activity
DNase-, RNase-, Protease-, Endotoxin- Free
TOCs
< 50 ppb
Resistivity
> 18 MΩ-cm
Treatment
Not DEPC-Treated
Quantity
100 mL, 500 mL, 1000 mL
Purification Methods
Activated charcoal, polyphosphate, reverse osmosis, electro-deionization, ion exchange, UV, ultrafiltration
Shipping Conditions
Room Temperature
Details
Applications For use in any molecular biology application
Storage Conditions Norgen’s Nuclease-Free Water should be stored at room temperature. Storage at +4°C or -20°C is optional.
Precautions and Disclaimers This product is designed for research purposes only. It is not intended for human or diagnostic use.
