Sputum Liquification Buffer (Cat. 28289) can be purchased separately
This kit constitutes an all-in-one system for the rapid isolation of DNA from sputum samples. It allows for the isolation of bacterial or eukaryotic DNA from the sputum samples using spin-column chromatography based on Norgen’s proprietary resin. The kit includes all the reagents needed to process sputum DNA. The protocol can be completed in 30 minutes. Purified DNA is of the highest quality and free from inhibitors, and can be used in sensitive downstream applications including PCR and qPCR.
Background
A sputum specimen is the name given to the mucus which is expectorated from the lower airways. High quality sputum samples should contain very little saliva, as this may contaminate the sputum sample with oral bacteria. Sputum samples are typically evaluated to look for infections such as Moraxella catarrhalis, Mycobacterium tuberculosis, Streptococcus pneumoniae ;and Haemophilus influenza. Other pathogens can be detected in sputum as well, including HIV. In addition, sputum samples are useful in the detection of lung cancer or to evaluate chronic inflammation.
* Average DNA yield will vary depending on the health status of the donor
Storage Conditions and Product Stability All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment.
Norgen’s Sputum DNA Isolation Kit contains ready-to-use Proteinase K, which is dissolved in a specially prepared storage buffer. The buffered Proteinase K is stable for up to 2 years after the date of shipment when stored at room temperature.
Component
Cat. 46200 (25 preps)
Slurry D
55 mL
Proteinase K in Storage Buffer
0.6 mL
Solution WN
4 mL
Wash Solution BE
9 mL
Elution Buffer B
8 mL
Mini Filter Spin Columns
25
Collection Tubes
25
Elution Tubes (1.7 mL)
25
Product Insert
1
Other Products
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.
Short term stability: 2-8oC, Long term stability: See individual component labels
Stability:
> 1 year under recommended storage conditions
Analyte:
D-Lactic Acid, L-Lactic Acid
Assay Format:
Spectrophotometer
Detection Method:
Absorbance
Wavelength (nm):
340
Signal Response:
Increase
Linear Range:
0.5 to 30 µg of D- or L-lactic acid per assay
Limit of Detection:
0.21 mg/L
Reaction Time (min):
~ 10 min (L-lactic acid), ~ 5 min (D-lactic acid)
Application examples:
Wine, soft drinks, milk, dairy products, foods containing milk (e.g. dietetic foods, bakery products, baby food, chocolate, sweets and ice-cream), vinegar, fruit and vegetables, processed fruit and vegetables, meat products, food additives, paper (and cardboard), cosmetics, pharmaceuticals and other materials (e.g. biological cultures, samples, etc.).
Method recognition:
Methods based on this principle have been accepted by DIN, GOST, IDF, EEC, EN, ISO, OIV, IFU, AIJN and MEBAK
The D-/L-Lactic Acid (D-/L-Lactate) (Rapid) test kit is used for the rapid and specific concurrent measurement and analysis of L-lactic acid (L-lactate) and D-lactic acid (D-lactate) in beverages, meat, dairy and food products.
Note for Content: The number of manual tests per kit can be doubled if all volumes are halved. This can be readily accommodated using the MegaQuantTM Wave Spectrophotometer (D-MQWAVE).
Extended cofactors stability. Dissolved cofactors stable for > 1 year at 4oC.
Rapid total analysis time (concurrent / flexible D and L-lactic acid reaction format)
D-lactate dehydrogenase reaction very rapid with most samples (~ 5 min)
Very competitive price (cost per test)
All reagents stable for > 2 years after preparation
Mega-Calc™ software tool is available from our website for hassle-free raw data processing
Standard included
Document
The D-/L-Lactic Acid (D-/L-Lactate) (Rapid) test kit is used for the rapid and specific concurrent measurement and analysis of L-lactic acid (L-lactate) and D-lactic acid (D-lactate) in beverages, meat, dairy and food products.
Isolate genomic DNA from animal tissues, cells, bodily fluids, viruses and swabs
Rapid and convenient spin column procedure
Purified DNA is of the highest quality and integrity for sensitive downstream applications including PCR, qPCR, genotyping, sequencing and more
This kit is designed for the rapid preparation of genomic DNA from various tissue samples, cultured cells, viruses, bodily fluids and swabs using a rapid spin column protocol. Purified DNA is of an excellent yield and quality, and is immediately ready for any downstream application including PCR, qPCR, genotyping, sequencing and more. The protocol can be completed in approximately 80 minutes (including incubation time).
Average Yield:* HeLa Cells (1 x 106) Tissue (from 10 mg kidney)
8 µg 10 µg
Maximum Amount of Starting Material: Animal Tissues Cultured Cells Bodily Fluids (blood, saliva) Viral Suspension
20 mg 3 x 106 cells 150 µL 150 µL
Time to Complete 10 Purifications
80 minutes
* Yield will vary depending on the type of sample processed
Storage Conditions and Product Stability The Proteinase K should be stored at -20°C upon arrival and after reconstitution. All other solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment.
