Lyophilized format designed for room temperature shipping
Available in TaqMan format for analysis
Norgen’s Vibrio cholerae TaqMan PCR Lyophilized Kit is designed for the detection of Vibrio cholerae specific DNA in a real-time PCR based on the use of TaqMan® technology. The lyophilized format is designed to ship the kit at ambient temperature.
Norgen’s Vibrio cholerae TaqMan Lyophilized Probe/Primer and Control Set is designed for the detection of Vibrio cholerae specific DNA in a real-time PCR based on the use of TaqMan® technology. The lyophilized format is designed to ship the kit at ambient temperature.
All kit components should be stored at -20°C upon arrival.
Once reconstituted, repeated thawing and freezing (>2 times) of the Master Mix and Positive Control should be avoided, as this may affect the performance of the assay. If the reagents are to be used only intermittently, they should be frozen in aliquots.
All kit components can be stored for 2 years after the date of production without showing any reduction in performance.
Component
Cat. TM38550L (100 preps)
Cat. TM38510L (100 preps)
MDx TaqMan 2X PCR Master Mix (Lyo)
4 x 350 μL
–
Vibrio cholerae Primer & Probe Mix (Lyo)
280 μL
280 μL
Vibrio cholerae Positive Control (Lyo)
150 μL
150 μL
Nuclease-Free Water (Negative Control)
3 x 1.25 mL
1 x 1.25 mL
Product Insert
1
1
Other Products
OT-2 Robot
Product Info
Document
Product Info
The Opentrons® OT-2 is a high-precision, open-source liquid handler designed to be easy for scientists to use and flexible enough to automate many application workflows. This robot and its accompanying automation platform are used to automate hundreds of protocols and workflows in genomics, proteomics, cell-based assays, and drug discovery.
The OT-2 has a user-friendly, plug and play graphical interface to design protocols compatible with any ANSI/SBS compliant or automation friendly labware. The base model includes an Opentrons GEN2 Single-Channel Pipette; you can also use up to two pipettes at the same time. Additional pipettes can be purchased separately and are easily interchangeable by the user. Consumables and other labware are sold separately.
Document
The Opentrons® OT-2 is a high-precision, open-source liquid handler designed to be easy for scientists to use and flexible enough to automate many application workflows. This robot and its accompanying automation platform are used to automate hundreds of protocols and workflows in genomics, proteomics, cell-based assays, and drug discovery.
The OT-2 has a user-friendly, plug and play graphical interface to design protocols compatible with any ANSI/SBS compliant or automation friendly labware. The base model includes an Opentrons GEN2 Single-Channel Pipette; you can also use up to two pipettes at the same time. Additional pipettes can be purchased separately and are easily interchangeable by the user. Consumables and other labware are sold separately.
Methylation Specific Bisulfite Seq Library Prep Kit
Product Info
Document
Product Info
Bisulfite seq is a well know technology to detect DNA methylation and several technologies such as WGBS, RRBS, MeDIP-Seq, and MSBS are used for whole genome DNA methylation analysis. DNA methylation is important for regulation of cell development, differentiation and gene expression in molecular biology, genetics and epigenetics. Most methylated cytosines are found at CpG sites, and 70-80% of cytosines are methylated. The number of CpG sites in human genome is around 28 million, which is less than 1% of the genome compared with 4.4% expected.
Whole genome bisulfite sequencing (WGBS) is the most effective method of DNA methylation analysis. The only limitation is the sequencing cost is very high because the whole genome is sequenced including all the non-methylated regions.
Reduced Representation Bisulfite Sequencing (RRBS) is the reduced representation of a smaller fraction of the methylated CpG sites. RRBS combines restriction enzyme digestion and bisulfite sequencing, and enriches the sequencing for methylated CpG sites. It is an efficient technology for estimate the whole genome methylation patterns at the single base level. Although this allows a higher coverage depth and reduces the sequencing cost, the limitation is only 10% of the methylated CpG sites are covered.
Methylated DNA Immunoprecipitation Sequencing (MeDIP-Seq) is another whole genome enrichment technique used for selection of methylated DNA. Using antibodies against 5-methylcytosine, methylated DNA is enriched from whole genomic DNA via immunoprecipitation. 5-methylcytosine antibodies are incubated with fragmented genomic DNA and precipitated, followed by DNA purification and sequencing. There are several drawbacks of MeDIP-Seq: 1. Low resolution (150~200 bp) as opposed to the single base resolution; 2. Non-specific interaction due to antibody specificity and selectivity. 3. Bias towards hypermethylated regions.
The Methylation Specific Bisulfite Seq (MSBS) Library Prep Kit (illumina platform) was developed for construction of NGS libraries for methylated CpG sites using bisulfite treated DNA (20 ng – 500 ng) as input. The kit enriches methylated CpG regions, thus significantly reduce the sequencing cost. The kit estimates the whole genome methylation patterns at the single base level since it is based on a bisulfite-seq technology.
It is known that bisulfite treatment of completed NGS libraries causes tremendous damage to the libraries. By using bisulfite treated DNA as input, the kit overcomes the significant library loss due to the bisulfite conversion. The kit contains a mixture of PCR polymerases that have high-fidelity amplification and uracil tolerance which is ideal for bisulfite treated DNA.
Methylation Specific Bisulfite Seq Library Prep Kit Workflow
Three index types are available for the kit:
Non-index (Cat.# 30101): Libraries do not have index.
Index (Cat.# 30102): 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.# 30103): 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.
Methylation Specific Bisulfite Seq advantages
Enrichment of methylated CpG sites
Single-base resolution
Low cost for sequencing
Fast
Total time: 1.5 hours
Hands-on time: 10 minutes
Simple workflow
Bisulfite treated DNA as input: From 20 ng to 500 ng
MSBS Library Prep Kit enriches CpG sites
High methylation regions and low methylation regions in human genome.
High methylation region in human genome.
Low methylation region in human genome.
Sequencing setting: Single-end 35 cycles (Read 1, 35 bases) recommended To maximize the methylated CpG enrichment, we recommend to sequence the MSBS libraries with single end 35 cycles (read1, 35 bases). This is because the enriched methylated CpG sites are mainly located around the beginning of read 1 sequences. Shorter single end reads tend to have better methylated CpG enrichment.
Document
Bisulfite seq is a well know technology to detect DNA methylation and several technologies such as WGBS, RRBS, MeDIP-Seq, and MSBS are used for whole genome DNA methylation analysis. DNA methylation is important for regulation of cell development, differentiation and gene expression in molecular biology, genetics and epigenetics. Most methylated cytosines are found at CpG sites, and 70-80% of cytosines are methylated. The number of CpG sites in human genome is around 28 million, which is less than 1% of the genome compared with 4.4% expected.
1.5 mL – 2 mL Micro-centrifuge Tube or Screw-Cap Vial Chiller Block
SKU: CB024
Permagen’s SBS/ANSI footprint 24-Tube thermal block allows the use of up to twenty four 1.5 mL -2 mL Microcentrifuge tubes, or screw-cap vials for keeping sample temperatures consistent. Can be used for heating as well
Document
Permagen’s SBS/ANSI footprint 24-Tube thermal block allows the use of up to twenty four 1.5 mL -2 mL Microcentrifuge tubes, or screw-cap vials for keeping sample temperatures consistent. Can be used for heating as well
