Opentrons Flex Filter Tips, 200 µL

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

• Detection kits for Streptococcus agalactiae
• Lyophilized format designed for room temperature shipping
• Available in TaqMan format for analysis

Norgen’s Streptococcus agalactiae TaqMan PCR Lyophilized Kit is designed for the detection of Streptococcus agalactiae specific DNA in a real-time PCR based on the use of TaqMan® technology. The lyophilized format is designed to ship the kit at ambient temperature.

Norgen’s Streptococcus agalactiae TaqMan Lyophilized Probe/Primer and Control Set is designed for the detection of Streptococcus agalactiae specific DNA in a real-time PCR based on the use of TaqMan® technology. The lyophilized format is designed to ship the kit at ambient temperature.

Details

Supporting Data

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Storage Conditions and Product Stability

• All kit components should be stored at -20°C upon arrival.
• Once reconstituted, repeated thawing and freezing (>2 times) of the Master Mix and Positive Control should be avoided, as this may affect the performance of the assay. If the reagents are to be used only intermittently, they should be frozen in aliquots.
• All kit components can be stored for 2 years after the date of production without showing any reduction in performance.

Component	Cat. TM30650L (100 preps)	Cat. TM30610L (100 preps)
MDx TaqMan 2X PCR Master Mix (Lyo)	4 x 350 μL	–
Streptococcus agalactiae Primer & Probe Mix (Lyo)	280 μL	280 μL
Streptococcus agalactiae Positive Control (Lyo)	150 μL	150 μL
Nuclease-Free Water (Negative Control)	3 x 1.25 mL	1 x 1.25 mL
Product Insert	1	1

Overview

• All-in-one solution for inclusion body protein isolation and purification
• Fast and convenient spin column protocol
• Complete kit with Cell Lysis Reagent, Inclusion Body Solubilization Reagent, buffers and spin columns to purify proteins
• Purification is based on spin column chromatography that uses Norgen’s resin separation matrix

These kits provide everything required to isolate and purify inclusion body proteins from induced bacterial cultures. First a proprietary Cell Lysis Reagent is used to selectively lyse the cells and release inclusion bodies in their solid form. Next, inclusion bodies are dissolved and their contents released using the provided IB Solubilization Reagent. Inclusion body proteins are then further purified using spin columns for rapid and convenient buffer exchange and desalting. This kit provides a convenient way to screen recombinants prior to scaling up.

ProteoSpin™ Inclusion Body Protein Isolation Micro Kit

The process is efficient and streamlined and can process up to 12 samples in only 60 minutes. Each spin column is able to recover up to 50 µg of acidic or basic proteins. Purified recombinant proteins are then ready for SDS-PAGE, 2D gels, Western blots, Mass Spectrometry analysis, and other applications.

ProteoSpin™ Inclusion Body Protein Isolation Maxi Kit

The procedure is efficient and streamlined and can process up to 4 samples in approximately 2 hours. Each spin column is able to recover up to 12 mg of acidic or basic proteins from 100 mL of induced bacterial culture. Purified recombinant proteins are then ready for SDS-PAGE, 2D gels, Western blots, Mass Spectrometry analysis, and other applications.

About Inclusion Bodies

Bacteria are widely used for the expression of different proteins. However, 70-80% of the proteins expressed in bacteria by recombinant techniques are typically contained in insoluble inclusion bodies (i.e., protein aggregates). The protein of interest found in these subcellular structures is often inactive, due to incorrect folding. The production rate of recombinant proteins stored in inclusion bodies is invariably higher than those synthesized as soluble proteins. The reason behind this is thought to be the resistance of insoluble proteins to proteolysis by cellular enzymes. In addition, separation of insoluble recombinant proteins in inclusion bodies is considerably easier than that of soluble proteins. These factors have been the major influences favoring scale-up of high-value proteins using bacterial fermentation for example. Procedures for the purification of the expressed proteins from inclusion bodies are often labour-intensive, time-consuming and not cost-effective. This kit provides the essential reagents for cell disruption, inclusion body solubilization and purification using spin column chromatography – all optimized to work together thereby simplifying the process and saving a tremendous amount of time and cost.

Details

Supporting Data

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Kit Specifications
Maximum Culture Volume	1.5 mL
Yield from 1.5 mL	Up to 50 μg
Minimum Elution Volume	30 μL
Time to Process 12 Samples	60 minutes

Storage Conditions

The Cell Lysis Reagent and IB Solubilization Reagent should be stored at 4°C upon receipt of this kit. All other solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment. Once opened, the solutions should be stored at 4°C when not in use except for the pH Binding Buffers (Acidic and Basic). Some precipitation will occur with 4°C storage. This precipitation should be dissolved with slight heating to room temperature before using.

Component	Cat. 10300 (Micro – 25 preps)	Cat. 17700 (Maxi – 4 preps)
Wash Solution C	30 mL	130 mL
Wash Solution N	30 mL	130 mL
Binding BUffer A	4 mL	20 mL
Binding Buffer N	4 mL	20 mL
Elution Buffer C	8 mL	2 x 30 mL
Protein Neutralizer	4 mL	4 mL
Cell Lysis Reagent	15 mL	110 mL
IB Solubilization Reagent	2 mL	50 mL
Syringes, 1cc, slip tip	25	–
Needles (Bev, 20G x 1 inch)	25	–
Syringes, 10 mL, Luer-Lok™ Tip	–	4
Needles (18G x 1.5 inch)	–	4
Micro Spin Columns	25	–
Maxi Spin Columns (filled with SiC) inserted into 50 mL collection tubes	–	4
Collection Tubes	25	–
Elution Tubes (1.7 mL)	25	–
Elution Tubes (50 mL)	–	4
Product Insert	1	1