t-Boc-N-Amido-PEG2-propargyl

The NGS Cell Free DNA Library Prep Kit (illumina and MGI Platforms) was developed for the construction of high quality cell-free DNA (cfDNA) libraries using 1 ng to 50 ng of cell-free DNA as input. The kit has a simple work flow and a fast procedure. Multiplexing of the cell free DNA library is possible based on the index type.

NGS Cell Free DNA Library Prep Kit Workflow

The main source of cell-free DNA (cfDNA) is derived from apoptotic hematopoietic cells in blood and found in the plasma. The length of the cfDNA is about 150-200 bp in length. Circulating tumor DNA (ctDNA) derived from malignant tumors is a part of cfDNA. Both cfDNA and ctDNA can be used as a noninvasive biomarker since it offers a better approach in comparison to invasive tissue biopsies.

NGS has been used for cfDNA and ctDNA sequencing in the field of liquid biopsy as it provides a whole genome level of molecular profiling. One of the hurdles for cfDNA sequencing is the difficulty of library preparation from the limited amount of cfDNA obtained from plasma. Our cell-free NGS kit makes it easy to get enough libraries from limited input in just 1.5 hours.

Three index types are available for the NGS Cell Free DNA Library Prep Kit of the illumina platform:

Non-index (Cat.# 30029): Libraries do not have index.

Index(Cat.# 30031): Each of our index primers contains a unique 6-base index sequence that can be used for sample identification. Total 48  library multiplexing is possible. Index information can be downloaded here.

Unique dual index (Cat.# 30033): Cell-free DNA library multiplexing up to 96 samples is possible with the unique dual indexes. We have developed a Four-Base Difference Index System. The system have at least 4 bases different from each other in the 8 bases index length. The primers effectively minimize sequencing errors such as mis-assignment, index hopping, index contamination etc. Index information can be downloaded here.

Indexes are available for the MGI platform kits (Cat.# 34031).

Kit advantages:

• Fast and simple protocol
  • Hands-on time is around 10 minutes
  • The total protocol time is only 1.5 hours
• Easy procedure based on:
  • Ready-to-use master mix for easy setup of reactions
  • Less reaction components to simplify bench work
• Less magnetic beads used for cleanup procedures: This can reduce more than 50% of the beads consumption
• Guaranteed high library conversion efficiency
• Low input cell-free DNA: From 1 ng to 50 ng

Comparison of library conversion efficiency with different samples under the same condition.

Comparison of library yield with different samples under the same condition.

The NGS Cell Free DNA Library Prep Kit (illumina and MGI Platforms) was developed for the construction of high quality cell-free DNA (cfDNA) libraries using 1 ng to 50 ng of cell-free DNA as input. The kit has a simple work flow and a fast procedure. Multiplexing of the cell free DNA library is possible based on the index type.

Overview

• Clean-up & concentrate total RNA (including miRNA) in minutes
• Clean-up & concentrate RNA from TRIzol^®, TRI Reagent^®, etc
• Clean-up RNA from contaminants including enzymes, primers, nucleotides
• Rapid spin-column protocol, elute in 20 µL
• 96-well format for high throughput processing & Micro-Elute spin column formats for 8 µL are available
• Purified RNA is suitable for a variety of downstream applications, including Small RNA Sequencing. Find out more information on Norgen’s NGS services
• Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix

These kits are designed to clean-up and concentrate RNA (including miRNA) from TRIzol^® and TRI Reagent^®, enzymatic reactions, in vitro transcription, labelling reactions, etc. These are robust kits for all clean-up and concentration purposes for up to 35 µg of RNA in solution. The purified RNA is of the highest purity and integrity, and can be used in a number of downstream applications. These kits purify all sizes of RNA, from large mRNA and ribosomal RNA down to miRNA, siRNA and lncRNA.

RNA Clean-Up and Concentration Kit (Spin Column)

Norgen’s RNA Clean-Up and Concentration Kit provides a rapid method for the purification, cleanup and concentration of up to 35 μg of RNA isolated using different methods including phenol/guanidine-based protocols, and from various upstream enzymatic reactions such as DNase treatment and labeling. The kit elutes RNA in 20 µL. Complete 10 purifications in 20 minutes.

RNA Clean-Up and Concentration 96-Well Kit (High Throughput)

Norgen’s RNA Clean-Up and Concentration 96-Well Kit provides a rapid method for the purification, cleanup and concentration of up to 50 μg of RNA isolated using different methods including phenol/guanidine-based protocols, and from various upstream enzymatic reactions such as DNase treatment and labeling. The kit elutes RNA in 75 µL. Complete 10 purifications in 30 minutes.

RNA Clean-Up and Concentration Micro-Elute Kit (Micro-Elute)

Norgen’s RNA Clean-Up and Concentration Micro-Elution Kit provides a rapid method for the purification, cleanup and concentration of up to 10 μg of RNA isolated using different methods including phenol/guanidine-based protocols, and from various upstream enzymatic reactions such as DNase treatment, labeling and in vitro transcription. This kit is made for eluting RNA in smaller volumes of 8 µL for all types of downstream applications, for the highest concentration of RNA sample. Complete 10 purifications in 20 minutes.

Protocol

Details

Supporting Data

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Kit Specifications (Spin Column)
Maximum Column Binding Capacity	35 μg
Size of RNA Purified	All sizes, including small RNA (<200 nt)
Maximum Amount of Starting Material	35 μg of RNA
Minimum Elution Volume	20 μL
Time to Complete 10 Purifications	20 minutes
Average Recovery	≥ 90%

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.

Component	Cat. 23600 (50 preps)	Cat. 43200 (100 preps)	Cat. 25100 (192 preps)	Cat. 61000 (50 preps)
Buffer RL	40 mL	2 x 40 mL	2 x 40 mL	40 mL
Wash Solution A	38 mL	2 x 38 mL	2 x 38 mL	38 mL
Elution Solution A	6 mL	2 x 6 mL	2 x 20 mL	6 mL
Column Activation Solution	–	–	–	30 mL
Micro Spin Columns	50	100	–	–
Micro-Elute RNA Spin Columns	–	–	–	50
96-Well Plate	–	–	2	–
Adhesive Tape	–	–	4	–
Collection Tubes	50	100	–	50
96-Well Collection Plate	–	–	2	–
Elution Tubes (1.7 mL)	50	100	–	50
96-Well Elution Plate	–	–	2	–
Product Insert	1	1	1	1