N-(t-butyl ester-PEG4)-N-bis(PEG4-Propargyl) is a trifunctional PEG linker that combines a t-butyl ester with two terminal alkynes. The alkynes can be applied in copper-click chemistry with azides to form stable triazole linkages with a target molecule, while the t-butyl ester can be deprotected and reacted with amines or alcohols to form amides or esters.
N-(t-butyl ester-PEG4)-N-bis(PEG4-Propargyl) is a trifunctional PEG linker that combines a t-butyl ester with two terminal alkynes. The alkynes can be applied in copper-click chemistry with azides to form stable triazole linkages with a target molecule, while the t-butyl ester can be deprotected and reacted with amines or alcohols to form amides or esters.
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Library size selection is an enrichment of a specific range of library sizes for NGS library preparations. 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, or SPRI (Solid Phase Reversible Immobilization) beads, is well used for the purification of DNA due to their reversible DNA binding. The NGS library can be size-selected by the magnetic beads or SPRI beads. The properties of the magnetic beads can be changed for a specific range of DNA binding. The contaminants and other unwanted components in the libraries can also be removed during size selection.
Specific ranges of NGS libraries can be selected using magnetic beads with different buffer compositions. The first DNA-beads binding step, also called the right-side clean-up, removes large NGS library fragments. The large NGS library fragments that bind to the beads are discarded with the beads pellet. The desired NGS library fragments in the supernatant are transferred to a new well, and new beads are added to the supernatant for the second beads-DNA binding, also called the left-side clean-up. After the rinsing step, the NGS library fragments with the dual selection are eluted in water or an appropriate buffer. The magnetic beads method has great advantages over time-consuming column purification and tedious gel-based purification.
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Library size selection for long-read sequencing only requires a single clean-up. In this case, only the large library fragments are bound to the beads, while other small library fragments are discarded with the supernatant. The selected larger library fragments are eluted in water or an appropriate buffer after the rinsing step.
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These kits provide fast and reliable procedures for the purification of genomic DNA from viable yeast cells, fungal spores or mycelium, and bacteria including Gram-positive. Genomic DNA is efficiently extracted from the cells by a combination of heat treatment, detergents and Bead Tubes (provided). An optional lyticase treatment allows for improved DNA yields with certain fungal and yeast species. Recovered genomic DNA is of excellent yield and purity for any downstream application including PCR, qPCR, Restriction Fragment Length Polymorphism (RFLP), Amplified Fragment Length Polymorphism (AFLP), sequencing, SNP analysis and more.
Fungi/Yeast Genomic DNA Isolation Kit (Spin Column)
This kit provides rapid spin column purification of genomic DNA from viable yeast cells, fungal spores or mycelium, and bacteria including Gram-positive. Preparation time for a single sample is less then 30 minutes, and each kit contains sufficient materials for 50 preparations.
Fungi/Yeast Genomic DNA Isolation 96-Well Kit (HT)
Norgen’s Fungi/Yeast Genomic DNA Isolation 96-Well Kit provides a fast, reliable and simple procedure for high throughput isolation of DNA from viable yeast cells, fungal spores or mycelium and Gram-positive bacteria. The purification could be performed on either a vacuum manifold or using centrifugation. Complete 96 purifications in 40 minutes.
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| Kit Specifications | |
| Column Binding Capacity | 50 μg |
| Maximum Column Loading Volume | 650 μL |
| Maximum Amount of Starting Material: Fungi (wet weight) Yeast or Bacteria Culture | 50 mg 0.5mL – 1mL |
| Average Yield* Pichia sp. (yeast) Botrytis cinerea Fusarium sp. Aspergillis niger Mucor racemosus Cladosporium cladosporioides Penicillium sp. Rhizopus oryzae Alternaria tenuissima | 25 μg 32 μg 42 μg 26 μg 15 μg 12 μg 40 μg 7 μg 30 μg |
| Time to Complete 10 purifications | 30 minutes |
* Yield will vary depending on the type of sample processed
Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 1 year after the date of shipment.
| Component | Cat. 27300 (50 preps) | Cat. 27350 (192 preps) |
|---|---|---|
| Lysis Buffer L | 30 mL | 2 x 60 mL |
| Resuspension Solution A | 20 mL | 60 mL |
| Solution BX | 28 mL | 2 x 28 mL |
| Wash Solution A | 18 mL | 2 x 38 mL |
| Elution Buffer B | 15 mL | 30 mL |
| Bead Tubes | 50 | 200 |
| Spin Columns | 50 | – |
| Collection Tubes | 50 | – |
| 96-Well Plate | – | 2 |
| 96-Well Collection Plate | – | 2 |
| Adhesive Tape | – | 4 |
| Elution Tubes (1.7 mL) | 50 | – |
| 96-Well Elution Plate | – | 2 |
| Product Insert | 1 | 1 |
