CO00019 Midstream Cassette 140x15mm, 86×4.5mm blue w/lens

Overview

• Convenient ready-to-use solution
• High sensitivity and yield
• Robust amplification

Norgen’s 2X PCR Master Mix is a ready-to-use solution that contains components required for PCR amplification including Taq DNA polymerase, dNTPs, reaction buffer, MgCl₂, KCl and a PCR enhancer/stabilizer. The user needs only to add template, the primer set and water to the master mix in order to set up the PCR reaction. This convenient 2X PCR Master Mix reduces the time required to set up PCR reactions and reduces the possibility of contamination, particularly when preparing large numbers of reactions. The optimized master mix allows for robust amplification of DNA templates with high yields of PCR products.

Taq DNA Polymerase is a highly thermostable DNA polymerase that possesses a 5´→ 3´ polymerase activity and a very low 5´→ 3´ exonuclease activity. The source of Taq included with Norgen’s 2X PCR Master Mix is an E. coli strain with a cloned Taq DNA Polymerase gene from Thermus aquaticus YT-1.

Details

Supporting Data

Figure 1 / 2

Previous

Next

Click for expanded view

Reagents Supplied – Cat # 28007

•  2X PCR Master Mix (1 Vial, 100 Reactions) – Sufficient reagent for 100 x 20 µL reactions

Storage Conditions and Product Stability
Norgen’s 2X Master Mix should be stored at -20ºC. For everyday use an aliquot can be stored at 4ºC for up to three months. The Master Mix is stable for multiple freeze-thaw cycles (see Figure 2). When stored at the proper temperature this reagent is stable for at least 1 year.

Description

The EzRNA™ T7 High Yield RNA Synthesis Kit is a user-friendly product for enzymatic RNA production. The enzyme mix contains adequate amount of T7 RNA polymerase, pyrophosphatase, and RNase inhibitors for in vitro transcription (IVT). Along with 10X Transcription Buffer and NTP Premix, users can swiftly assemble IVT reactions without compromising RNA yield. The EzRNA™ T7 High Yield RNA Synthesis Kit allows for the attainment of approximately up to 150 µg RNA yield within 2 hours at 37°C.

Features

• High yield
• Versatile- suitable for short and long transcripts
• NTP premixed- Minimal pipetting and setup time
• Compatible with CleanCap® Reagent AG
• Lithium chloride included for RNA purification

Application

• Generation of RNA from T7 promoter-driven DNA sequences
• Suitable for subsequent cap-0 and cap-1 modification

Storage

-20°C for 12 months

The EzRNA™ T7 High Yield RNA Synthesis Kit is a user-friendly product for enzymatic RNA production. The enzyme mix contains adequate amount of T7 RNA polymerase, pyrophosphatase, and RNase inhibitors for in vitro transcription (IVT). Along with 10X Transcription Buffer and NTP Premix, users can swiftly assemble IVT reactions without compromising RNA yield. The EzRNA™ T7 High Yield RNA Synthesis Kit allows for the attainment of approximately up to 150 µg RNA yield within 2 hours at 37°C.

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.