HiPure FFPE RNA Kit supplies a simple and rapid RNA extraction for Formalin-fixed, paraffin-embedded (FFPE) tissue and sections samples. This kit is based on silica gel column purification technology, no phenol-chloroform extraction or alcohol precipitation. The whole extraction only takes 30 minutes (not including digestion time). RNA can be directly used for downstream applications such as RT-PCR, Northern blot, vitro translation and other experiments.
Detail
Introduction
HiPure FFPE RNA Kit supplies a simple and rapid RNA extraction for Formalin-fixed, paraffin-embedded (FFPE) tissue and sections samples. This kit is based on silica gel column purification technology, no phenol-chloroform extraction or alcohol precipitation. The whole extraction only takes 30 minutes (not including digestion time). RNA can be directly used for downstream applications such as RT-PCR, Northern blot, vitro translation and other experiments.
Details
Specifications
Features
Specifications
Main Functions
Isolation total RNA from FFPE tissue and section samples
Applications
RT-PCR, quantitative RT-PCR, Northern hybridization, Poly A purification, nucleic acid protection and in vitro translation
Purification method
Mini spin column
Purification technology
Silica technology
Process method
Manual (centrifugation or vacuum)
Sample type
FFPE tissue sample
Sample amount
6mg
Yield
20μg
Elution volume
≥20μl
Time per run
≤60 minutes
Liquid carrying volume per column
800µl
Binding yield of column
100µg
This product is based on silica column purification. Remove paraffin by Buffer DPS. Sample lysis with proteinase K digestion requires only 15 minutes. After lysis, samples are incubated at 80ºC for 15 minutes. Transfer to an adsorption column and RNA is adsorbed on the membrane, while protein is not adsorbed and is removed with filtration. After washing proteins and other impurities, RNA was finally eluted with low-salt buffer.
Advantages
High quality – high purity total RNA can be directly used in various sensitive downstream applications
Fast – several samples can be extracted in 60 minutes by column method
Safe – no phenol chloroform extraction required
Sensitive – RNA can be recovered at the level of PG
Kit Contents
Contents
R414302
D414303
Purification Times
50 Preps
250 Preps
HiPure RNA Micro Columns
50
250
2ml Collection Tubes
50
250
Buffer DPS
60 ml
250 ml
Buffer FRL
15 ml
60 ml
Buffer RLC
15 ml
60 ml
Buffer RWC*
10 ml
50 ml
Buffer RW2*
20 ml
2 x 50 ml
Protease Dissolve Buffer
1.8 ml
10 ml
Proteinase K
24 mg
120 mg
RNase Free Water
10 ml
20 ml
Storage and Stability
Proteinase K should be stored at 2-8°C upon arrival. However, short-term storage (up to 12 weeks at room temperature (15-25°C) does not affect their performance. The remaining kit components can be stored at room temperature (15-25°C) and are stable for at least 18 months under these conditions. The entire kit can be stored at 2-8°C, but in this case buffers should be redissolved before use. Make sure that all buffers are at room temperature when used.
Other Products
Sircol-2.0 Soluble Collagen assay kit
Product Info
Document
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.
Document
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.
NGS DNA Fragmentation & Library Prep Kit (illumina and MGI Platforms)
Product Info
Document
Product Info
The NGS DNA Fragmentation & Library Prep Kit (illumina and MGI Platforms) was developed for construction of high quality libraries for next generation sequencing. The kit uses intact genomic DNA (EDTA-free DNA or DNA resuspended in TE buffer) as input DNA without an additional DNA fragmentation step. Our technology provides a fast and simple workflow. DNA libraries can be generated around 2 hours with only 10 min hands-on time. Library multiplexing is possible.
NGS DNA Fragmentation & Library Prep Kit Workflow
The incorporation of DNA fragmentation in the kit makes it possible to directly use intact genomic DNA as input DNA without the need of mechanical DNA shearing or enzymatic DNA fragmentation. The NGS DNA Fragmentation & Library Prep Kit does not generate sequencing bias as compared to library using mechanical sheared DNA as input. Sequence coverage is also consistent between enzymatic shearing and mechanical shearing. The library size is inversely correlated with the incubation time of step 1 at 20°C.
Three index types are available for the kit of the illumina platform:
Non-index (Cat.# 30026): Libraries do not have index.
Index (Cat.# 30028): Each of the index primers contains a unique index sequence of 6 bases. Library multiplexing for 48 samples is possible. Index information can be downloaded here.
Unique dual index (Cat.# 30030): Library multiplexing for 96 samples is possible. With the unique features of our 4-Base Difference Index System, the index sequence is 8-base long and each index has 4 bases different from others. Our unique dual index primers effectively identify sequencing errors such as index hopping, mis-assignment of reads, and de-multiplexing errors etc. The unique dual index primers set consists of 96 pre-mixed unique pairs of index primers. Index information can be downloaded here.
Indexes are available for the MGI platform kits (Cat.# 34028).
Kit features:
1.5-hour protocol from intact genomic DNA to NGS library
Intact genomic DNA as input, DNA fragmentation is not needed.
Works with both EDTA-free DNA and DNA resuspended in TE buffer
Library conversion efficiency: 100 ng, 300 ng and 500 ng of intact genomic DNA were used as input.
The library size is inversely correlated with the incubation time of step 1 at 20°C.
NGS data comparison: enzymatic shearing versus mechanical shearing
Enzymatic shearing • DNA shearing and library prep: BioDynami NGS DNA Fragmentation & Library Prep Kit Mechanical shearing • DNA shearing: Covaris sonication • Library prep: BioDynami NGS DNA Library Prep Kit.
Document
The NGS DNA Fragmentation & Library Prep Kit (illumina and MGI Platforms) was developed for construction of high quality libraries for next generation sequencing. The kit uses intact genomic DNA (EDTA-free DNA or DNA resuspended in TE buffer) as input DNA without an additional DNA fragmentation step. Our technology provides a fast and simple workflow. DNA libraries can be generated around 2 hours with only 10 min hands-on time. Library multiplexing is possible.