The PCR-free NGS DNA Library Prep Kit was developed for construction of DNA libraries for next generation sequencing (illumina platform). The kit adds 3′-dT-tailed library adapters to both ends of DNA fragments efficiently. The kit uses double strand DNA fragments (blunt and/or sticky) as input DNA for NGS library construction, and is compatible with DNA fragments generated from both enzymatic methods (BioDynami DNA fragmentation enzymes etc.) and physical methods (sonication, nebulization etc.).
Detail
The PCR-free NGS DNA Library Prep Kit was developed for construction of DNA libraries for next generation sequencing (illumina platform). The kit adds 3′-dT-tailed library adapters to both ends of DNA fragments efficiently. The kit uses double strand DNA fragments (blunt and/or sticky) as input DNA for NGS library construction, and is compatible with DNA fragments generated from both enzymatic methods (BioDynami DNA fragmentation enzymes etc.) and physical methods (sonication, nebulization etc.).
PCR-Free NGS DNA Library Prep Kit Workflow
PCR amplification is a standard step for library preparation of Next-Generation Sequencing. The PCR is used to amplify fully ligated DNA fragments and to add index information to the libraries. The indexing is for the pooling of the library samples in an effort to reduce the sequencing cost.
However, PCR introduces uneven amplification of DNA in some DNA regions with extreme GC-contents and secondary structures. This bias can cause very low sequencing coverage in such regions. DNA sequencing in these regions is still a huge challenge.
PCR-free library prep can reduce library bias and minimize sequencing gaps. The sequencing data from PCR-free library samples have even genomic coverage with few gaps and better depth in GC-rich regions. Our PCR-free NGS kit offers optimal coverage in the regions that are traditionally difficult, such as high-GC content regions, low-GC content regions, and repetitive sequence regions.
Two index types are available for the kit:
Non-index: Libraries do not have index.
Index: Each library contains one i5 index and one i7 index. Library multiplexing up to 96 samples is possible. List of indexes can be downloaded Here.
Kit advantages:
Total time: 1 hr
Hands-on time: 5 min
Easy procedure: Ready-to-use master mix & Less reaction components
Input DNA amount from 100 ng to 1 ug
Other Products
Sircol-2.0 Soluble Collagen assay kit
Product Info
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Product Info
What is Collagen?
Collagen is a fundamental component of the extracellular matrix, and the predominant protein in animals, constituting around 30% of total protein mass. A glycoprotein, it is well known for its triple helical structure. This is formed from three polypeptide α-chains with Gly-X-Y repeating residues (Gly for Glycine, X for proline, and Y for hydroxyproline).
Types of Collagen
Over 28 types of collagens have been identified, with Type I collagen being the most abundant. It’s prevalent in ligaments, tendons, skin, and bone tissue. Its mature, insoluble form grants it remarkable strength, making it vital for the mobility of organisms. Collagen also has biochemical functions, influencing cell growth, proliferation, and differentiation.
This version of the kit is designed to detect and measure SOLUBLE forms of collagen. Chose the Sircol Insoluble collagen kit if you need to analyse INSOLUBLE collagen.
Applications of Collagen
Collagen, with its diverse properties, finds utility in various industries. It plays a role in medicine for wound healing and has an expanding role in tissue engineering and cell culture for biomedical purposes. It’s gaining popularity in the cosmetic industry for skin rejuvenation and is used in chemical formulations and the food industry as a functional food supplement and additive.
How does Sircol 2.0 detect collagen?
The Sircol 2.0 dye reagent includes Sirius Red, a linear anionic dye with sulfonic acid side chains. This reagent is specially formulated to bind to the Gly-X-Yn helical structure of soluble collagen under assay conditions.
*The improved formulation of Sircol 2.0 dye enables a greater degree of dye-collagen specificity (compared to our previous S1000 assay kit).
Overview of the Sircol 2.0 assay process:
Step 1. Prepared samples are placed in the wells of the assay microplate, together with Sircol Dye Reagent. After 30 minutes mixing, any collagen-dye complexes will form as a precipitate. These are collected on the base of the microplate wells by centrifugation.
Step 2. Unbound dye is removed by gentle aspiration, followed by a rinse with Plate Wash Reagent.
Step 3. Following further centrifugation, collagen-bound dye is eluted by incubation with a Dye Release Reagent. Eluted dye is detected ‘in-situ’ by spectrophotometric analysis of the microplate at 556nm.
Step 4. The collagen content of unknown samples can be quantified by comparison against a calibration curve, prepared using the Collagen Reference Standard supplied with the kit.
A list of suggested sample types can be found under the ‘Assay Specification‘ tab.
Colorimetric Detection (556nm) (Endpoint), Requires a microplate centrifuge.
Measurements per kit
96 in total (allows a maximum of 41 samples to be run in duplicate alongside a standard curve).
Suitable Samples
Soluble* collagens from mammalian**:
In-vivo: Tissues, cartilages and fluids.
In-vitro: Extracellular matrices / Conditioned media from 2D/3D culture environments.
The straightforward sample processing and analysis of Sirco 2.0 make it a good alternative to conventional hydroxyproline analysis.
*Prior salt/acid/acid-pepsin extraction may be necessary to release soluble collagen.
**Sircol 2.0 is primarily designed for use with in-vivo / in-vitro samples of mammalian origin. Collagens originating from other taxonomic groups and kingdoms can also be analysed. See note on p6 of manual for further information.
Precautions
This kit is designed for research use only. Not for use in diagnostic procedures. Kit requires access to a microplate centrifuge* (see note below), as well as a spectrophotometer/colorimeter capable of absorbance detection at 556nm. Specific sample preparation protocols may require customer to provide further reagents, consult assay manual for further information.
*As a minimum, we recommend that the centrifuge can centrifuge a 96-well microplate at 400 x g for 120 minutes. Higher speed centrifuges are recommended (up to a maximum of 2000 x g), allowing a reduction in centrifuge time.
Sircol 2.0 kit contents:
1. Dye Reagent (1x20ml)
2. Collagen Reference Standard (1x5ml, 200µg/ml of soluble Bovine collagen)
3. Plate Wash Reagent (1x28ml)
4. Collagen Concentration Reagent (1x25ml)
5. Neutralisation Reagent (1x8ml)
6. Dye Release Reagent (1x25ml)
7. Assay Microplate (1×96-wells)
8. Microplate Seals (6x)
9. Documentation (QuickStart Guide / Manual / Certificate of Analysis)
NB: Additional reagents may be required for sample preparation prior to assay. Consult manual or contact us for further details. This kit requires the use of a microplate centrifuge, capable of centrifuging a 96-well microplate at 400 x g for 120 minutes. Higher speed centrifuges are recommended (up to a maximum of 2000 x g), allowing a reduction in centrifuge time.
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Experience user-friendly detection & measurement of Soluble Collagen with Sircol™ 2.0! Our latest kit simplifies collagen quantification within in-vivo / in-vitro samples. Sircol 2.0 offers enhanced sensitivity and accuracy compared to our previous Sircol kit.
Round well with ‘V’ bottom for maximum liquid extraction
High uniformity from well to well
Raised well rims for reliable closing with heat sealing.
Easy and reliable stacking
Good centrifugation stability up to 6,000 × g for faster protocols and improved sample quality
Manufactured under DNase/ RNase free environment without slip agents, plasticizers or biocides – Materials which could have a negative effect on bioassays
Pure, virgin polypropylene guarantees good extractible performance, high resistance to chemicals, good mechanical stress and working with temperature extremes
Autoclavable (121 °C, 20 min)
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Working volume of 0.36ml
Round well with ‘V’ bottom for maximum liquid extraction
High uniformity from well to well
Raised well rims for reliable closing with heat sealing.
Easy and reliable stacking
Good centrifugation stability up to 6,000 × g for faster protocols and improved sample quality
Manufactured under DNase/ RNase free environment without slip agents, plasticizers or biocides – Materials which could have a negative effect on bioassays
Pure, virgin polypropylene guarantees good extractible performance, high resistance to chemicals, good mechanical stress and working with temperature extremes
Autoclavable (121 °C, 20 min)
[TP1000] ExcelTaq™ Taq DNA Polymerase, 5 U/μl, 500 U
Product Info
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Product Info
Description
ExcelTaq™ Taq DNA Polymerase is a recombinant thermo-stable Taq DNA polymerase expressed and purified from an E. coli strain carrying the cloned gene. With a high DNA synthesis rate and high thermo-stability, ExcelTaq™ Taq DNA Polymerase is suitable for common and specialized PCR applications.
Features
5’→3′ DNA polymerase activity
5’→3′ exonuclease activity
No detectable 3’→5′ exonuclease (proofreading) activity
Generates PCR products with 3’-dA overhangs
Thermo-stable – half-life lasts for more than 40 min at 95°C
Applications
Routine PCR
Amplification of DNA fragments up to 8 kb
Generation of PCR products for TA cloning
DNA labeling
Storage
-20°C for 24 months
Document
ExcelTaq™ Taq DNA Polymerase is a recombinant thermo-stable Taq DNA polymerase expressed and purified from an E. coli strain carrying the cloned gene. With a high DNA synthesis rate and high thermo-stability, ExcelTaq™ Taq DNA Polymerase is suitable for common and specialized PCR applications.
