Propargyl-PEG4-sulfonic acid

Description 

Champion™ E. coli Transformation Kit provides an easy method for rapid preparation of chemically competent cells with high transformation efficiency from fresh culture, overnight culture, or even directly from bacterial colonies on the plate. The competent cell preparation method eliminates the requirement of time-wasting wash step. In addition, preparation of competent cells from overnight culture or directly from bacterial colonies provides flexibility to cloning experiment. The resultant competent cells can be immediately used or stored at -70°C for one year. This kit includes a specialized SMO-Broth™ medium and a unique Champion™ CC Buffer for culturing and preparing competent cells efficiently. Following the simple and quick competent cell preparation protocol from fresh culture, the transformation efficiency is typically ranged from 10⁸–10⁹ cfu/μg transformants/μg of pUC19 plasmid DNA, but varies depending on the E. coli strains. The resultant competent cells can be further transformed using time-saving transformation protocol, eliminating the requirement of heat-shock and recovery steps. 

Features

• Flexible– fresh culture, overnight culture, 4°C stored liquid culture or even colonies on agar plate can be used for transformation. 
• Fast and Easy– only few steps for preparation; suitable for time-saving transformation 
• High efficiency– up to 109 cfu/μg 
• Personalization– suitable for most E. coli strains

Kit Contents

					Component	Volume
					Champion™ CC Buffer	20 ml
					SMO-Broth™	100 ml x 2
					pUC19 Control Plasmid (10-4 μg/μl)	5 µl
					Instruction Manual	1
					Champion™ Competent Cell Preparation Card	1

Storage

4°C for 12 months

Champion™ E. coli Transformation Kit provides an easy method for rapid preparation of chemically competent cells with high transformation efficiency from fresh culture, overnight culture, or even directly from bacterial colonies on the plate. The competent cell preparation method eliminates the requirement of time-wasting wash step. In addition, preparation of competent cells from overnight culture or directly from bacterial colonies provides flexibility to cloning experiment. The resultant competent cells can be immediately used or stored at -70°C for one year. This kit includes a specialized SMO-Broth™ medium and a unique Champion™ CC Buffer for culturing and preparing competent cells efficiently. Following the simple and quick competent cell preparation protocol from fresh culture, the transformation efficiency is typically ranged from 108–109 cfu/μg transformants/μg of pUC19 plasmid DNA, but varies depending on the E. coli strains. The resultant competent cells can be further transformed using time-saving transformation protocol, eliminating the requirement of heat-shock and recovery steps. 

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.

Introducing the Fastin Assay Kit: Your Straightforward Solution for Elastin Quantification! Our user-friendly kit utilizes a dye-based method to measure elastin from in-vivo and in-vitro sources. It can be used to quantify various elastin forms, spanning from immature tropoelastin to mature, ‘insoluble’ elastin fibers.

Colorimetric Detection (513nm) (Endpoint)

Understanding Elastin: The Key to Tissue Flexibility

Tissues like lungs and arteries must maintain the ability to stretch and recoil repeatedly throughout an organism’s life. Elastin, a mature protein, is responsible for this elasticity and is usually present as insoluble fibers within the ECM. During development, these fibers are initially formed from a soluble precursor called tropoelastin.

‍What is the Fastin Assay?

The Biocolor Fastin assay is a user-friendly, dye-based means of quantifying elastins derived from both in-vivo and in-vitro sources. A variety of elastin forms can be assayed, from immature tropoelastin to mature ‘insoluble’ elastin fibres.

Further information on how the assay works can be found on the ‘Mode of Action‘ tab.

A list of suggested sample types can be found under the ‘Assay Specification‘ tab.

How does the Fastin assay detect Elastin?

The Fastin Dye Reagent contains an elastin-binding synthetic porphyrin, TPPS (5,10,15,20-tetraphenyl- 21H,23H-porphine). This affinity of TPPS for elastin was first observed when used as a ‘vital stain’ on live animals. Most tissues took up the dye initially but only elastin retained the TPPS molecules over time. [Winkelman, J. (1962), Cancer Res. 22, 589-596; Winkelman, J & Spicer, S. (1962), Stain Technol. 37, 303-305].

It has been proposed that the elastin binding of TPPS may be due to the retention of the acidic dye (which contains four charged sulfate groups) by the basic amino acid side chain residues of elastin.

‍How does the Fastin assay work?

Step 1. Incubation of samples containing soluble elastin with the Fastin Dye Reagent causes an elastin-dye complex to form. This insoluble complex then precipiates.

Step 2. Dye-labelled elastin is then isolated by centrifugation and the unbound dye removed. Elastin-bound dye is then eluted and measured spectrophotometrically.

Step 3. The elastin content of unknown samples can be calculated by comparison against a calibration curve prepared using a standard comprising water-soluble elastin (supplied with the kit).

Assay range

50 – 500µg/ml

Limit of Detection

50µg/ml

Detection Method

Colorimetric Detection (513nm) (Endpoint)

Measurements per kit

110 in total (allows a maximum of 48 samples to be run in duplicate alongside a standard curve).

Suitable Samples

In-vivo: tissues and fluids. Insoluble elastin will first require conversion to water soluble α-elastin using the oxalic acid reagents and extraction protocol supplied with the kit.  

In-vitro: Elastin produced by cells during 2D/3D cell culture. NB elastin in conditioned cell media is typically below the detection limit of the kit.

‍

Precautions

This kit is designed for research use only. Not for use in diagnostic procedures.
Kit requires access to a centrifuge, heated water bath or block, as well as a spectrophotometer or colorimeter capable of absorbance detection at 513nm.
Specific sample preparation protocols may require customer to provide further reagents, consult assay manual for further information.

Fastin elastin kit contents:‍

1. Dye Reagent (1x110ml)

2. α-elastin Reference Standard (1x5ml, 1.0 mg/ml soluble Bovine elastin)

3. Oxalic Acid (1x20ml) Precipitating Reagent (1x30ml)

4. Dye Dissociation Reagent (1x30ml)

5. Precipitating Reagent (1x30ml)

6. 1.5ml micro-centrifuge tubes for dye-labelling reaction.

7. Assay kit manual

NB: Additional reagents may be required for sample preparation prior to assay. Consult manual or contact us for further details.

Introducing the Fastin Assay Kit: Your Straightforward Solution for Elastin Quantification! Our user-friendly kit utilizes a dye-based method to measure elastin from in-vivo and in-vitro sources. It can be used to quantify various elastin forms, spanning from immature tropoelastin to mature, ‘insoluble’ elastin fibers.
Colorimetric Detection (513nm) (Endpoint)