resDNASEQ E.coli Residual DNA Quantitation kit (Without DNA Control)
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Note: Price not include shipment & duty, contact us to get full quote.
The resDNASEQ E.coli Residual DNA Quantitation kit is designed for the quantification of residual DNA from E. coli, in cell lines which are used for production of biopharmaceutical products. The resDNASEQ E.coli Residual DNA Quantitation kit use TaqManTM quantitative PCR to perform rapid, specifc quantitation of femtogram levels of residual host-cell or plasmid DNA. The kit was developed to meet the sensitivity requirements defined by WHO (10 ng E. coli DNA per therapeutic dose).
Detail
Description
Features of the sDNASEQ E.coli Residual DNA
Quantitation kit include:
Simpler and Rapid
Only three steps will be need for Sample Preparation and All components of the Sample Preparation Kit can be stored at room temperature.
Only one Reagent for qPCR;
Only 1.5 hours will be needed for the whole test.
Accurate
Perfect amplification curve, good amplification efficiency and good precision.
Highly sensitive quantitation using proven TaqMan™ real-time qPCR technology.
Limit of Detection (LOD): 1 pg/mL; Limit of Quantification (LOQ): 5 pg/mL
The recovery rate of different concentration samples in the linear range is between 70% and 130%
Kit Performance
Fig 1. Only three steps will be need for Sample Preparation and only 20 minitutes will be taken for Sample Preparation.
Fig 2. Seven concentration samples of 5fg/μL, 10fg/μL, 20fg/μL, 30fg/μL, 300fg/μL, 3pg/μL, 30pg/μL, 300pg/μL were detected. CV of each concentration was < 30%, Regression coefficient associated with standard solutions was 0.99975, and amplification efficiency was 100.068%.
Fig 3. Four concentration samples of 5fg/μL, 10fg/μL, 20fg/μL, and 30fg/μL were detected, and 10 multiple holes were detected for each concentration. The detection values of 5fg/μL and above were CV ＜30%.
Fig 4. DNA recovery can be determined by including samples spiked with known DNA amounts which are prepared from the corresponding DNA standards. Typically, the range for this value varies from 70% to 130%.
Fig 5. Only one Reagent for qPCR MIX.
Other Products
2X One-Step RT-PCR Master Mix
Product Info
Document
Product Info
Overview
Convenient ready-to-use solution
High sensitivity and yield
Robust amplification
Available in 3 convenient sizes: 100, 200 and 500 reactions (20 µL vol)
Norgen’s 2X One-Step RT-PCR Master Mix is a ready-to-use solution that contains components required for RT-PCR amplification of RNA templates. The mix includes M-MuLV reverse transcriptase, Taq DNA polymerase, dNTPs, reaction buffer, MgCl2, KCl, and a PCR enhancer/stabilizer. The user needs only to add the template, the primer set and water to the Master Mix to set up the RT-PCR reaction. This convenient 2X One-Step RT-PCR Master Mix reduces the time required to set up PCR reactions and reduces the possibility of contamination, particularly when preparing large numbers of reactions. The optimized master mix allows for robust amplification of RNA templates with high yields of PCR products.
Taq DNA Polymerase is a highly thermostable DNA polymerase that possesses a 5´→ 3´ polymerase activity and a very low 5´→ 3´ exonuclease activity. The source of Taq included with Norgen’s 2X One-Step RT-PCR Master Mix is an E. coli strain with a cloned Taq DNA Polymerase gene from Thermus aquaticus YT-1. M-MuLV Reverse Transcriptase is an RNA-directed DNA polymerase that can synthesize a complementary DNA strand initiating from a primer using either RNA (cDNA synthesis) or single-stranded DNA as a template. The source of the Reverse Transcriptase included with Norgen’s 2X One-Step RT-PCR Master Mix is an E. coli strain with a cloned reverse transcriptase gene from M-MuLV.
Norgen’s 2X One-Step RT-PCR Master Mix is available in 3 convenient sizes:
Cat # 28113 – 100 reactions (sufficient for 100 reactions x 20 µL reaction volume) Cat # 28114 – 200 reactions (sufficient for 200 reactions x 20 µL reaction volume) Cat # 28115 – 500 reactions (sufficient for 500 reactions x 20 µL reaction volume)
2X One-Step RT-PCR Master Mix (1 Vials, 100 Reactions) – Sufficient reagent for 100 x 20 µL reactions
Storage Conditions and Product Stability 2X One-Step RT-PCR Master Mix should be stored at -20°C. For everyday use an aliquot can be stored at 4°C for up to 3 months. Repeated freeze-thaw cycles are not recommended. When stored at the proper temperature this reagent is stable for at least 1 year.
The series of DNA Size Selection Kits (Magnetic Beads) were developed for DNA size selection using magnetic beads. A total of 11 kits are available, with different selection ranges spanning from 50 bp to over 10 kb. The kits provide a simple and quick approach for the enrichment of a specific range of DNA fragments. The kit workflow allows double-sided or single-sided size selection for specific size cutoffs.
Gel images of different ranges of size selection. Sheared human genomic DNA was used as input.
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DNA size selection is a selective capture of DNA fragments of a specific range of size for next-generation sequencing (NGS) library preparations, PCR, ChIP assay, DNA ligations, endonuclease digestions, adapter removal, and other genomics and molecular biology applications. DNA size selection is preferred after NGS library prep in most of the cases. The NGS library preparation is related to the quality of the sequencing data. Precise NGS library size selection can increase sequencing efficiency, improve data quality, and reduce costs.
There are two types of sequencing technologies: short-read sequencing and long-read sequencing. Short-read sequencing uses DNA libraries that contain small insert DNA fragments of similar sizes, usually several hundred base pairs. The sequencing efficiency can be improved if the DNA size selection is in the right range. Cat.# 20104S and 20104L are the best kits for NGS library size selection of illumina paired-end 100 (PE100) sequencing with 100-200 bp library inserts; Cat.# 20105S and 20105L are the best kits for NGS library size selection of illumina paired-end 150 (PE150) sequencing with 150-300 bp library inserts; and Cat.# 20106S and 20106L are the best kits for NGS library size selection of illumina paired-end 300 (PE300) sequencing with 300-600 bp library inserts.
Long-read sequencing uses a large DNA fragment as input and makes very long reads. Usually, library size selection is preferred to remove smaller fragments. Cat.# 20110S and 20110L are the best kits for long-read sequencing size selection with DNA sizes >5 kb, and Cat.# 20111S and 20111L are the best kits for long-read sequencing size selection with DNA sizes >10 kb.
The magnetic beads technology uses paramagnetic particles, also known as SPRI (Solid Phase Reversible Immobilization) beads, to bind DNA reversibly and selectively. DNA fragments can be size-selected and purified by changing the properties of the magnetic beads or SPRI beads. The magnetic beads can easily separate the beads-binding DNA from the contaminants and unwanted components in the samples. The samples after DNA size selection are free of contaminants such as buffer components, enzymes, proteins, salts, dNTPs, primers, and adapters. Our proprietary magnetic beads reagents improve yield, selectivity, and reproducibility.
Specific DNA fragments at a certain length range can be purified simply using magnetic separation with different beads components, avoiding tedious and time-consuming gel extraction and column-based purification. The magnetic beads method is popular for common DNA size selection, including library size selection. The first beads-binding step, referred to as the right-side clean-up, removes large DNA fragments. The large DNA fragments are bound to the beads and are discarded. The desired DNA fragments in the supernatant are transferred to a new well, and new beads are added to the supernatant for the second beads-binding, referred to as the left-side clean-up. The double-size selected DNA fragments are eluted after ethanol rinsing.
DNA size selection with dual clean-ups.
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A single clean-up is needed for DNA size selection with large fragments. In this case, only the large DNA fragments are bound to the beads. The selected larger DNA fragments are eluted after ethanol rinsing.
DNA size selection with single clean-up for >5 kb and >10 kb DNA.
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Features of DNA size selection and library size selection
High specificity and high recovery of size selection
11 selection ranges are available, including 5 ranges for NGS library size selection
50-100 bp
100-200 bp
200-500 bp
250-350 bp: ideal for illumina PE100 sequencing
300-450 bp: ideal for illumina PE150 sequencing
450-750 bp: ideal for illumina PE300 sequencing
500-1000 bp
1-3 kb
1-5 kb
>5 kb: ideal for long-read sequencing
>10 kb: ideal for long-read sequencing
Fast and simple
20-min protocol
No gel purification required
No columns required
No centrifugation required
Efficient removal of contaminants and unwanted components
