DBCO-C5-PEG12-acid is an analog of DBCO-Acid with PEG linker and a DBCO group. DBCO group is a highly reactive cycloalkyne which is commonly used for copper-free Click Chemistry reactions. The hydrophilic PEG chain allows for increased water solubility of compounds in aqueous media. The terminal carboxylic acid can react with primary amine groups in the presence of activators (e.g. HATU) to form a stable amide bond. Reagent grade, for research use only.

DBCO-C5-PEG12-acid is an analog of DBCO-Acid with PEG linker and a DBCO group. DBCO group is a highly reactive cycloalkyne which is commonly used for copper-free Click Chemistry reactions. The hydrophilic PEG chain allows for increased water solubility of compounds in aqueous media. The terminal carboxylic acid can react with primary amine groups in the presence of activators (e.g. HATU) to form a stable amide bond. Reagent grade, for research use only.

The kit was developed for construction of high quality libraries for next generation sequencing (illumina and MGI Platforms). The kit needs double strand DNA fragments (blunt and/or sticky) as input DNA for NGS library preparation, and is compatible with DNA fragments generated from both enzymatic methods and physical methods (sonication, nebulization etc.). Library multiplexing is possible with different types of indexes.

The kit was optimized for next generation sequencing (NGS) library preparation with different types of samples. Most of DNA library preparation requires the ligation of sheared DNA fragments to library adaptors and the DNA library preparation is closely related to the quality of NGS data. With BioDynami’s unique DNA library preparation technologies, the fast and simple kit allows high quality NGS library preparation to be completed in 1.5 hours with only 10 minutes of hands-on time.

<img decoding="async" src="https://biodynami.com/wp-content/uploads/2020/07/NGS-DNA-Library-Prep-Kit-illumina-1.png" alt=""

Some genomic regions are very difficult to be covered evenly and usually result in very low coverage rate or gap in these regions.

Typical difficult regions are:
• with high GC contents
• have secondary structures: mainly due to repeat sequences
• the worst cases: have both high GC contents and repeated sequences.

Example: human TERT gene is one of the most difficult regions as shown above. NGS data showed that BioDynami kit has the best performance to cover the extremely difficult human TERT gene region.

Limited GC bias across whole genome

<img decoding="async" src="https://biodynami.com/wp-content/uploads/2020/07/BioDynami-NGS-DNA-Library-Prep-Kit-GC-bias.png" alt="

Three index types are available for the illumina platform kits:

Non-index (illumina Cat.# 30009): Libraries do not have index.

Index (illumina Cat.# 30021): A unique barcode sequence with 6 bases has been included in each of the index primers. RNA Sequencing library multiplexing is possible with up to 48 samples. Index information can be downloaded here.

Unique dual index (illumina Cat.# 30023): RNA Sequencing library multiplexing up to 96 samples is possible with the unique dual indexes. We have developed a 4-Base Difference Index System. The system can generate indexes with at least 4 bases different from others in the 8-base indexing region. the unique dual indexing primers identify sequencing errors such as index hopping, mis-assignment, and de-multiplexing errors. Index information can be downloaded here.

Indexes are available for the MGI platform kits (Cat.# 34021).

The kit was developed for construction of high quality libraries for next generation sequencing (illumina and MGI Platforms). The kit needs double strand DNA fragments (blunt and/or sticky) as input DNA for NGS library preparation, and is compatible with DNA fragments generated from both enzymatic methods and physical methods (sonication, nebulization etc.). Library multiplexing is possible with different types of indexes.

Introduction

This kit provides fast purification of high-quality RNA from cells, tissues, and yeast using silica-membrane spin columns with a binding capacity of 100μg RNA. There is no need for phenol/chloroform extractions and time-consuming steps such as CsCl gradient ultracentrifugation or isopropanol precipitation. RNA purified using the HiPure Total RNA Purification System can be used for applications such as RT-PCR, Northern blotting, poly A+ RNA (mRNA) purification, nuclease protection, and in vitro translation.

Details

Specifications

Features	Specifications
Main Functions	Isolation total RNA (not include miRNA) from 20mg tissue, 150mg plant, 5 x 106 cell using two columns (gDNA removed column)
Applications	RT-PCR, qRT-PCR, Northern hybridization, second generation sequencing
Purification method	Mini spin column
Purification technology	Silica technology, DNA filtration technology
Process method	Manual (centrifugation or vacuum)
Sample type	Animal soft tissue, cultured cells, lymphocytes, simple plant tissue
Sample amount	Cells:≤1 x 107 Animal tissue sample: 1-20 mgPlant tissue: 50-150 mg
Yield	2-100μg
Elution volume	≥50μl
Time per run	≤25 minutes（1-24 samples）
Liquid carrying volume per column	800µl
Binding yield of column	100µg

Principle

The Kit isolates total RNA from up to 10⁷ cells or 20 mg tissue. A short workflow enables RNA isolation with genomic DNA removal in less than 25 mins. Samples are first lysed and homogenized. The lysate is passed through a DNA Mini column, ethanol is added to the flow-through, and the sample is applied to a RNA column. RNA binds to the membrane and contaminants are washed away. High-quality RNA is eluted in as little as 30µl water using the Kit.

Advantages

• Efficient removal of DNA – unique genomic DNA removal column without DNase treatment
• High quality – high purity total RNA can be directly used in various sensitive downstream applications
• Fast – several samples can be extracted in 25 minutes by column method
• Safe – no phenol chloroform extraction required
• Sensitive – RNA can be recovered at the level of PG

Kit Contents

Contents	R411102	D411103
Purification Times	50 Preps	250 Preps
HiPure DNA Mini Columns	50	250
HiPure RNA Mini Columns	50	250
2ml Collection Tubes	100	2 x 250
Buffer RLC	50 ml	200 ml
Buffer RW1	50 ml	200 ml
Buffer RW2*	12 ml	2 x 50 ml
RNase Free Water	10 ml	30 ml

Storage and Stability

HiPureKit can be stored dry at room temperature (15-25°C) and are stable for at least18 months under these conditions. During shipment, crystals or precipitationmay form in the Buffer RLC. Dissolve by warming buffer to 37°C.

This kit provides fast purification of high-quality RNA from cells, tissues, and yeast using silica-membrane spin columns with a binding capacity of 100μg RNA. There is no need for phenol/chloroform extractions and time-consuming steps such as CsCl gradient ultracentrifugation or isopropanol precipitation. RNA purified using the HiPure Total RNA Purification System can be used for applications such as RT-PCR, Northern blotting, poly A+ RNA (mRNA) purification, nuclease protection, and in vitro translation.