Norgen’s FFPE RNA Purification Kits provide a rapid method for the isolation and purification of total RNA (including microRNA) from formalin-fixed paraffin-embedded (FFPE) tissue samples in as little as 1 hour. Using formalin to fix tissues leads to crosslinking of the RNA and proteins, and the process of embedding the tissue samples can also lead to fragmentation of the RNA over time. Norgen’s FFPE RNA Purification Kits provide conditions that allow for the partial reversing of the formalin modifications, resulting in a high quality and yield of RNA. These kits are able to purify all sizes of RNA, from large mRNA and ribosomal RNA down to microRNA (miRNA) and small interfering RNA (siRNA), depending on the age of the FFPE tissue as fragmentation of the RNA is known to occur over time. The RNA is preferentially purified from other cellular components without the use of phenol or chloroform.
FFPE RNA Purification Kit (Spin Column)
Maximum loading volume of 650 μL per column, and a maximum binding capacity of 50 μg per column.
FFPE RNA Purification 96-Well Kit (High Throughput)
Purification is based on 96-well column chromatography using Norgen’s proprietary resin as the separation matrix. Purification can be performed using either a vacuum manifold or centrifugation. Maximum loading volume of 400 μL per well, and a maximum binding capacity of 50 μg per well.
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| Kit Specifications | |
| Maximum Binding Capacity Per Well | 50 μg |
| Maximum Loading Volume Per Well | 400 μL |
| Size of RNA Purified | All sizes, including small RNA (< 200 nt) |
| Maximum Amount of Starting Material: | 5 slices of < 20 µm thick paraffin 25 mg of unsectioned block |
Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. 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. This kit is stable for 1 year after the date of shipment.
| Component | Cat. 25300 (50 preps) | Cat. 25400 (2 x 96 preps) |
|---|---|---|
| Digestion Buffer A | 25 mL | 2 x 25 mL |
| Buffer RL | 30 mL | 2 x 30 mL |
| Enzyme Incubation Buffer | 6 mL | 2 x 6 mL |
| Wash Solution A | 38 mL | 2 x 38 mL |
| Elution Solution A | 6 mL | 2 x 20 mL |
| Proteinase K | 12 mg | 2 x 20 mg |
| DNase I | 1 vial | 2 x 500μL |
| Micro Spin Columns | 50 | – |
| 96-Well Incubation Plate | – | 2 |
| 96-Well Plate | – | 2 |
| Adhesive Tape | – | 8 |
| Collection Tubes | 50 | – |
| 96-Well Collection Plate | – | 2 |
| Elution Tubes (1.7 mL) | 50 | – |
| 96-Well Elution Plate | – | 2 |
| Product Insert | 1 | 1 |
The Bisulfite Sequencing Library Prep Kit (illumina platform) was developed for construction of NGS libraries using bisulfite treated DNA (50 ng – 500 ng) as input. DNA methylation is an epigenetic mechanism known to play a critical role in gene regulation and genomic imprinting by blocking transcription factor access to promoters and enhancers. Bisulfite sequencing is a popular technique in biomedical research based on C to T conversion under the treatment of sodium bisulfite.
Recently, NGS became a powerful tool to identify the DNA methylation status at the whole genome level with single-base resolution. However, it is well known that bisulfite treatment of the NGS libraries causes tremendous damage to the libraries.
Bisulfite-Seq kit comparison
In the case of the regular bisulfite seq library preparation (library prep before bisulfite conversion), the DNA shearing equipment and the expensive methylated adaptors are required. In addition, the subsequent bisulfite conversion causes tremendous DNA damage to the constructed libraries.
BioDynami has developed a unique library prep technology to solve the problems. The technology uses bisulfite treated DNA as input to avoid the significant library loss caused by bisulfite conversion. Furthermore, DNA shearing step and expensive methylated adaptors are not required with our kit. The DNA polymerase in the kit has high-fidelity amplification ability and uracil tolerance which is ideal for amplification of bisulfite sequencing libraries. The final library is strand specific.
Bisulfite Sequencing Library Prep Kit Workflow
Three index types are available for the kit:
Non-index (Cat.# 30091): Libraries do not have index.
Index (Cat.# 30092): 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.# 30093): 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.
Kit advantages
The Bisulfite Sequencing Library Prep Kit (illumina platform) was developed for construction of NGS libraries using bisulfite treated DNA (50 ng – 500 ng) as input. DNA methylation is an epigenetic mechanism known to play a critical role in gene regulation and genomic imprinting by blocking transcription factor access to promoters and enhancers. Bisulfite sequencing is a popular technique in biomedical research based on C to T conversion under the treatment of sodium bisulfite.
Nucleic acid testing (NAT) is the method of choice for detection and quantification of a wide range of micro organisms. Primerdesign manufactures and supplies high quality quantitative real-time PCR kits for the detection and simultaneous quantification of numerous significant pathogens . A copy number standard curve is provided for quantification and an the internal extraction template (DNA or RNA), controls for the quality of the nucleic acid extraction and eliminates false negative results.
The kit is designed with the broadest possible detection profile to ensure that all clinically relevant strains and subtypes are detected. Target sequences are selected by working with data from key opinion leaders in the field. Multiple sequence alignments and unprecedented real-time PCR expertise in design and validation ensure the best possible kit.
Details of the target and priming specificity are included in the individual handbooks above.
Packaged, optimised and ready to use. Expect Better Data.
Exceptional value for money
Rapid detection of all clinically relevant subtypes
Positive copy number standard curve for quantification
Highly specific detection profile
High priming efficiency
Broad dynamic detection range (>6 logs)
Sensitive to < 100 copies of target
Accurate controls to confirm findings