GeneAb™ MUC6

Product Description

High-Efficiency Isothermal RNA Amplification Kit, Stable And Easy To Operate

Product Detail

Kit Storage and term of Validity

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

This kit is based on a rapid nucleic acid amplification technology at room temperature and constant temperature:at room temperature and constant temperature(generally 39℃~42℃),reverse transcriptaseuses specific primers and template RNA tosynthesize cDNA strands,and binds the auxiliary protein and single strand with the help of the protein, the recombinase and the primer form a complex; perform a homology search and bind the target homology domain, at this time a D-loop region is formed at the homology positionand strand exchange begins; accompanied by the recombinase from the complex upon dissociation,the polymerase also bindsto the 3′ end of the primer,initiating chainextension.It is suitable for laboratory-level RNA amplification and RNA amplification for other detection purposes.

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.

Technical Parameters:

Parameters	Details
Product Name	RNA Isothermal Amplification Kit Basic
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 Applications

Suitable for RNA isothermal rapid amplification kit(Basic type)

Primer: Require pair of nucleotide primers with the length of 25-35 bp.

RNA 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.

The series of DNA Size Selection Kits (Magnetic Beads) were developed for DNA size selection using magnetic beads. A total of 11 kits are available, with different selection ranges spanning from 50 bp to over 10 kb. The kits provide a simple and quick approach for the enrichment of a specific range of DNA fragments. The kit workflow allows double-sided or single-sided size selection for specific size cutoffs.

Gel images of different ranges of size selection. Sheared human genomic DNA was used as input.

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DNA size selection is a selective capture of DNA fragments of a specific range of size for next-generation sequencing (NGS) library preparations, PCR, ChIP assay, DNA ligations, endonuclease digestions, adapter removal, and other genomics and molecular biology applications. DNA size selection is preferred after NGS library prep in most of the cases. The NGS library preparation is related to the quality of the sequencing data. Precise NGS library size selection can increase sequencing efficiency, improve data quality, and reduce costs.

There are two types of sequencing technologies: short-read sequencing and long-read sequencing. Short-read sequencing uses DNA libraries that contain small insert DNA fragments of similar sizes, usually several hundred base pairs. The sequencing efficiency can be improved if the DNA size selection is in the right range. Cat.# 20104S and 20104L are the best kits for NGS library size selection of illumina paired-end 100 (PE100) sequencing with 100-200 bp library inserts; Cat.# 20105S and 20105L are the best kits for NGS library size selection of illumina paired-end 150 (PE150) sequencing with 150-300 bp library inserts; and Cat.# 20106S and 20106L are the best kits for NGS library size selection of illumina paired-end 300 (PE300) sequencing with 300-600 bp library inserts.

Long-read sequencing uses a large DNA fragment as input and makes very long reads. Usually, library size selection is preferred to remove smaller fragments. Cat.# 20110S and 20110L are the best kits for long-read sequencing size selection with DNA sizes >5 kb, and Cat.# 20111S and 20111L are the best kits for long-read sequencing size selection with DNA sizes >10 kb.

The magnetic beads technology uses paramagnetic particles, also known as SPRI (Solid Phase Reversible Immobilization) beads, to bind DNA reversibly and selectively. DNA fragments can be size-selected and purified by changing the properties of the magnetic beads or SPRI beads. The magnetic beads can easily separate the beads-binding DNA from the contaminants and unwanted components in the samples. The samples after DNA size selection are free of contaminants such as buffer components, enzymes, proteins, salts, dNTPs, primers, and adapters. Our proprietary magnetic beads reagents improve yield, selectivity, and reproducibility.

Specific DNA fragments at a certain length range can be purified simply using magnetic separation with different beads components, avoiding tedious and time-consuming gel extraction and column-based purification. The magnetic beads method is popular for common DNA size selection, including library size selection. The first beads-binding step, referred to as the right-side clean-up, removes large DNA fragments. The large DNA fragments are bound to the beads and are discarded. The desired DNA fragments in the supernatant are transferred to a new well, and new beads are added to the supernatant for the second beads-binding, referred to as the left-side clean-up. The double-size selected DNA fragments are eluted after ethanol rinsing.

DNA size selection with dual clean-ups.

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A single clean-up is needed for DNA size selection with large fragments. In this case, only the large DNA fragments are bound to the beads. The selected larger DNA fragments are eluted after ethanol rinsing.

DNA size selection with single clean-up for >5 kb and >10 kb DNA.

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Features of DNA size selection and library size selection

• • • High specificity and high recovery of size selection
    • 11 selection ranges are available, including 5 ranges for NGS library size selection
      • • 50-100 bp
        • 100-200 bp
        • 200-500 bp
        • 250-350 bp: ideal for illumina PE100 sequencing
        • 300-450 bp: ideal for illumina PE150 sequencing
        • 450-750 bp: ideal for illumina PE300 sequencing
        • 500-1000 bp
        • 1-3 kb
        • 1-5 kb
        • >5 kb: ideal for long-read sequencing
        • >10 kb: ideal for long-read sequencing
    • Fast and simple
      • • 20-min protocol
        • No gel purification required
        • No columns required
        • No centrifugation required
    • Efficient removal of contaminants and unwanted components

Overview

• Extract high quality & quantity total RNA including miRNA 
• Isolate from a wide variety of animal tissues 
• No phenol step required – isolate all RNA in one fraction
• Compatible with tissues stored in RNAlater® or Trizol®  
• Convenient & fast spin column format
• Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix

This kit is suitable for the isolation of total RNA from a range of animal tissues such as liver and spleen, as well as difficult fibrous tissues such as heart, muscle, intestine, etc.  Briefly, the tissue of interest is first lysed using Buffer RL, followed by treatment with the provided Proteinase K which aids in the removal of the various proteins present in fiber-rich tissues including collagen, contractile proteins, and connective tissues. The purified RNA is of the highest quality and purity, with excellent RIN values and A260/A280, and is suitable for downstream applications including qRT-PCR, RT-PCR, microarrays, NGS, and more.

The kit purifies all sizes of RNA from large mRNA, lncRNA down to microRNA (miRNA) in the same fraction without the requirement of phenol. Isolate all RNA sequences at an equal rate irrespective of size. Moreover, when the RNA sequences are small (e.g. miRNA), the column binds small RNAs regardless of their GC content.

Details

Supporting Data

Figure 1 / 4

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Kit Specifications
Maximum Column Binding Capacity	50 μg
Maximum Column Loading Volume	650 μL
Size of RNA Purified	All sizes, including small RNA < 200 nt
Time to Complete 10 Purifications	50 minutes
Average Yield Liver (10mg) Kidney (10mg) Spleen (10mg) Heart (10mg) Muscle (10mg)	30 μg 25 μg 15 μg 10 μg 5 μg
Maximum Amount of Starting Material    Heart Kidney Liver Muscle  Spleen	30 mg 15 mg 15 mg 30 mg 15 mg

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

Component	Cat. 25700 (50 preps)
Buffer RL	30 mL
RNase-Free Water	40 mL
Wash Solution A	38 mL
Enzyme Incubation Buffer A	6 mL
Elution Solution A	6 mL
Proteinase K	2 vials
DNase I	1 vial
Mini Spin Columns	50
Collection Tubes	100
Elution Tubes (1.7 mL)	50
Product Insert	1