The ExcelRT™ One-Step RT-PCR Kit is designed for the reverse transcription and PCR amplification of a specific target RNA from either total RNA or mRNA. The ExcelRT™ One-Step RT-PCR Kit provides the user an alternative to the lengthy two step processes (first strand generation and amplification) by using a single mixture, single tube, one step reaction. The ExcelRT™ One-Step RT-PCR Kit contains a 2X reaction premix consisting of an optimized buffer, dNTPs, Mg2+ and enzyme stabilizer, and a blend of recombinant reverse transcriptase and Taq DNA polymerase. The ExcelRT™ One-Step RT-PCR Kit allows the user to complete the RT-PCR process using a thermocycler in a single reaction setting. The ExcelRT™ One-Step RT-PCR Kit is capable of detecting even trace amounts of target RNA and ideal for target RNA amplification and analysis.
Detail
Description
The ExcelRT™ One-Step RT-PCR Kit is designed for the reverse transcription and PCR amplification of a specific target RNA from either total RNA or mRNA. The ExcelRT™ One-Step RT-PCR Kit provides the user an alternative to the lengthy two step processes (first strand generation and amplification) by using a single mixture, single tube, one step reaction. The ExcelRT™ One-Step RT-PCR Kit contains a 2X reaction premix consisting of an optimized buffer, dNTPs, Mg2+ and enzyme stabilizer, and a blend of recombinant reverse transcriptase and Taq DNA polymerase. The ExcelRT™ One-Step RT-PCR Kit allows the user to complete the RT-PCR process using a thermocycler in a single reaction setting. The ExcelRT™ One-Step RT-PCR Kit is capable of detecting even trace amounts of target RNA and ideal for target RNA amplification and analysis.
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
Round well with ‘U’ bottom to aid mixing, pelleting and washing
High uniformity from well to well
Raised well rims for reliable closing with heat sealing.
Easy and reliable stacking
Good centrifugation stability up to 6,000 × g for faster protocols and improved sample quality
Manufactured under DNase/ RNase free environment without slip agents, plasticizers or biocides – Materials which could have a negative effect on bioassays
Pure, virgin polypropylene guarantees good extractible performance, high resistance to chemicals, good mechanical stress and working with temperature extremes
Autoclavable (121 °C, 20 min)
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Working Volume of 2.0ml Round well with ‘U’ bottom to aid mixing, pelleting and washing High uniformity from well to well Raised well rims for reliable closing with heat sealing. Easy and reliable stacking Good centrifugation stability up to 6,000 × g for faster protocols and improved sample quality Manufactured under DNase/ RNase free environment without slip agents, plasticizers or biocides – Materials which could have a negative effect on bioassays Pure, virgin polypropylene guarantees good extractible performance, high resistance to chemicals, good mechanical stress and working with temperature extremes Autoclavable (121 °C, 20 min)
