The RNA Seq Library Prep Kit was developed for construction of high quality NGS libraries for next generation sequencing (illumina and MGI Platforms). RNA Sequencing is a very powerful tool to analyze transcriptome such as gene expression and transcription regulation, splicing characterization, mutation and variation detection etc. The kit needs purified RNA (example: rRNA depleted RNA or polyA mRNA) as input for library construction. Library multiplexing is possible with different types of indexes.
Three index types are available for the illumina platform kits:
Non-index (illumina Cat.# 30055): Libraries do not have index.
Index (illumina Cat.# 30056): A unique barcode sequence with 6 bases has been included in each of the index primers. RNA Sequencing library multiplexing is possible with up to 48 samples. Index information can be downloaded here.
Unique dual index (illumina Cat.# 30057): RNA Sequencing library multiplexing up to 96 samples is possible with the unique dual indexes. We have developed a 4-Base Difference Index System. The system can generate indexes with at least 4 bases different from others in the 8-base indexing region. the unique dual indexing primers identify sequencing errors such as index hopping, mis-assignment, and de-multiplexing errors. Index information can be downloaded here.
Indexes are available for the MGI platform kits (Cat.# 34112).
Features
Magnetic Beads (tRNA Purification)
Solid Phase Reversible Immobilization magnetic beads has been used for nucleic acids purification because they are effective, simple, and fast. The carboxyl groups coated beads are paramagnetic particles that reversibly bind to nucleic acid. However, there is a drawback for magnetic beads that it can only purify DNA/RNA that are 100 base pairs or longer. tRNA and other DNA/RNA fragments shorter than 100 base pairs are not effectively recovered. We have developed Magnetic Beads (tRNA Purification) to overcome the hurdle.
The Magnetic Beads (tRNA Purification) solves the tRNA and short DNA/RNA purification problem. The beads with our proprietary technology purify tRNA effectively by removing impurities and unwanted components such as dNTPs, detergents, salts, proteins, and other contaminants. The magnetic bead reagents can be used for oligo (>70 nt) applications.
In the case of samples contaminated with RNA/DNA such as rRNA and DNA, our magnetic beads can effectively remove RNA/DNA that are 180 nt and larger. Purified tRNA are ideal for applications requiring high quality, as the fragments are free of impurities and contaminants.
Comparison of tRNA and 70 nt oligos recovery. BioDynami beads (tRNA purification) successfully recovered DNA fragments both yeast tRNA and 70 nt oligos.
Recovery of tRNA and 70 nt oligos with BioDynami magnetic Beads (tRNA Purification). Yeast tRNA and 70 nt oligos were used as input. Input and recovered oligos were quantified with ssDNA Quantification kit (BioDynami Cat. # 40043) and RNA Quantification kit (BioDynami Cat. # 40044).
Depletion of larger RNAs. Left panel: depletion of 28S rRNA and 18S rRNA. Right panel: depletion of RNA of 180 nt and larger.
Features
Removal of unwanted components and other impurities
Purification of tRNA and oligos (>70 nt)
tRNA
RNA fragments 70 nt or longer
DNA/RNA hybrid fragments 70 nt or longer
Oligo and chimeric oligo 70 nt or longer
dsDNA fragments 70 bp or longer
ssDNA fragments 70 nt or longer
Removal of larger RNA/DNA contamination:
18S rRNA
28S rRNA
RNA/DNA> 180 nt
Solid Phase Reversible Immobilization magnetic beads has been used for nucleic acids purification because they are effective, simple, and fast. The carboxyl groups coated beads are paramagnetic particles that reversibly bind to nucleic acid. However, there is a drawback for magnetic beads that it can only purify DNA/RNA that are 100 base pairs or longer. tRNA and other DNA/RNA fragments shorter than 100 base pairs are not effectively recovered. We have developed Magnetic Beads (tRNA Purification) to overcome the hurdle.
Product Details
Kit Size:
48 Reactions
Product Name:
DNA Isothermal Rapid Amplification Kit Fluorescent Type
Kit Type:
DNA
Reaction Volume:
50 μL
Storage Temperature:
-20℃
Application:
Nucleic Acid Amplification
Type:
Fluorescent Type
Reaction Time:
20mins
High Light:
,
,
Payment & Shipping Terms
Minimum Order Quantity
48T
Price
3.8$/T
Packaging Details
16T/bag,48T/box
Delivery Time
6days
Payment Terms
T/T,Paypal
Supply Ability
100000T/Month
Product Description
Product parameters:
Reagent component ( WLE8202KIT ,16T/bags,48T/Box ) | |||
Component | Specification | Quantity | Function |
A buffer | 1.6ml | 1 Tube | Buffer system mainly for stabilizing protein/enzyme and performance |
B buffer | 0.15ml | 1 Tube | Mainly activated systems such as magnesium ions |
Positive control template | 0.1ml | 1 Tube | Mainly the positive plasmid template is used to test the effectiveness of the kit |
Positive control primer mix | 0.06ml | 1 Tube | Mainly the primer combination of the positive control template |
Reagent Guide Manua | 16T/bags,48T/Box | 3 bags | Reagent technology of protein/enzyme system: freeze-dried powder, freeze-dried microspheres |
Principle overview
This kit is based on a room temperature and constant temperature nucleic acid rapid amplification technology: at room temperature and constant temperature, the recombinase and primer form the protein/single-stranded nucleotide complex Rec/ssDNA, with the help of auxiliary proteins and single-stranded binding protein SSB , invade the double-stranded DNA template; form a D-loop region at the invasion site, and start scanning the DNA double-strands; after finding the target region complementary to the primer, the Rec/ssDNA complex disintegrates, and the polymerase also binds to The 3′ end of the primer initiates chain extension. This kit relies on the action of exonuclease at 39 ºC, adding specific molecular probes designed based on the template, and using fluorescence monitoring equipment to achieve real-time monitoring of the amplification process of the target fragment.
Primer design
It is recommended to use primers with a length of 30-35 bp. Primers that are too short will affect amplification speed and detection sensitivity; primers are designed to avoid the formation of secondary structures that affect amplification; the amplicon length is recommended to be 150-300 bp, usually no more than 500 bp.
Fluorescent probe design
The probe sequence does not overlap with the specific primer recognition site, is 46-52 nt in length, and the sequence avoids palindromic sequences, internal secondary structures, and continuous repeated bases. The probe has four modification sites: the middle position ≥ 35 nt from the 5′ end is labeled with a dSpacer (tetrahydrofuran, THF) as the recognition site for exonuclease; the upstream of the THF site is labeled with a fluorescent group, and the downstream Label a quenching group, the distance between the two groups is 2-4 nt; THF is ≥15 nt from the 3′ end, and the 3′ end is labeled with a modifying group, such as an amine group, a phosphate group or a C3-Spacer.
Product features and advantages:
This kit has the advantages of high sensitivity, strong specificity,and short reaction time (only 20 minutes), and the reaction groups are in dry powder state, which is easy to operate and easy to store.
It can be applied to various brands of fluorescence quantitative PCR instruments, constant temperature fluorescence amplification instruments and other fluorescence detection equipment.
This kit is based on a room temperature and constant temperature nucleic acid rapid amplification technology: at room temperature and constant temperature, the recombinase and primer form the protein/single-stranded nucleotide complex Rec/ssDNA, with the help of auxiliary proteins and single-stranded binding protein SSB , invade the double-stranded DNA template; form a D-loop region at the invasion site, and start scanning the DNA double-strands; after finding the target region complementary to the primer, the Rec/ssDNA complex disintegrates, and the polymerase also binds to The 3′ end of the primer initiates chain extension. This kit relies on the action of exonuclease at 39 ºC, adding specific molecular probes designed based on the template, and using fluorescence monitoring equipment to achieve real-time monitoring of the amplification process of the target fragment.
83, On-nut 88/2 Prawet Sub-district, Prawet District, Bangkok, 10250, Thailand
Tel : 081-875-1869 , 02-328-7179
Email : [email protected]
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