Biotin-PEG4-alkyne is a PEG linker containing a biotin group and a terminal alkyne. Biotin is useful for affinity-based applications such as pull-down assays or for ligating with streptavidin proteins while terminal alkynes are used in copper (I) click chemistry with azides to form stable triazoles with a target molecule. The inclusion of a PEG linker in this molecule improves its aqueous solubility.

Biotin-PEG4-alkyne is a PEG linker containing a biotin group and a terminal alkyne. Biotin is useful for affinity-based applications such as pull-down assays or for ligating with streptavidin proteins while terminal alkynes are used in copper (I) click chemistry with azides to form stable triazoles with a target molecule. The inclusion of a PEG linker in this molecule improves its aqueous solubility.

Introduction

The Kit is designed for purification of total RNA, including miRNA and other small RNA molecules (18nt), from cultured cells and various animal and human tissues, including difficult-to-lyse tissues samples. Alternatively, a miRNA-enriched fraction and a total RNA (>200nt) fraction can be purified separately. This Kit combines phenol/guanidine-based lysis of samples and silica membrane–based purification of total RNA. MagZol Reagent, included in the kits, is a monophasic solution of phenol and guanidine thiocyanate, designed to facilitate lysis of tissues, to inhibit RNases, and also to remove most of the cellular DNA and proteins from the lysate by organic extraction.

Details

Specifications

Features	Specifications
Main Functions	Isolation miRNA and other small RNA molecules(18nt), from cultured cells and various animal and human tissues, using MagZol reagent and column
Applications	RT-PCR, Northern Blot, poly A+purification, nucleic acid protection and in vitro translation
Purification method	Mini spin column
Purification technology	Silica technology
Process method	Manual (centrifugation or vacuum)
Sample type	Animal tissues, adherent cells, suspension cells, bacteria, etc
Sample amount	Eukaryotic culture cells: ≤ 10^7, Animal tissue:<100mg, Yeast culture cells:<5 x10^7, Bacteria:<10^9
Elution volume	≥15μl
Time per run	≤40 minutes
Liquid carrying volume per column	800µl
Binding yield of column	100µg

Principle

Hipure silica gel column is based on glass fiber filter membrane with high binding force. Under the condition of high concentration of ionizing agent (such asguanidine hydrochloride or guanidine isothiocyanate), the filter membrane can adsorb nucleic acid through hydrogen bond and electrostatic, while protein andother impurities are not adsorbed and removed. The filter membrane adsorbed with nucleic acid is washed to remove the residual protein and salt. Finally, the nucleic acid adsorbed on the filter membrane can be washed out with low salt buffer (such as buffer TE) or water. The obtained nucleic acid has high purity and can be directly used in various downstream experiments.

This kit combines acid/guanidine (MagZol) extraction technology with glass fiber filter membrane purification, which can improve the extraction effect of complex samples and samples with low RNA content. After the sample is treated with MagZol reagent and chloroform, the supernatant is added with ethanol to provide appropriate binding conditions, then transferred to the purification column and centrifuged. Macromolecular RNA can be efficiently bound to the membrane. Collect the filtrate containing small RNA, add more ethanol to adjust the binding capacity of small RNA, the pollutants can be efficiently washed away by second cleaning. Finally, the purified RNA was eluted by RNase free water.

Advantages

• High quality – one step RNA extraction reagent combined with silica gel column can obtain the highest concentration
• Fast – several samples can be extracted in 40 minutes
• High applicability – samples including animals, plants, bacteria, cells, etc.
• High concentration – efficiently remove macromolecular RNA, enrich small RNA and improve sensitivity

Kit Contents

Contents	R431002	R431003
Purification Times	50 Preps	250 Preps
HiPure RNA Mini Columns	100	2 x 250
2ml Collection Tubes	100	2 x 250
MagZol Reagent	60 ml	270 ml
Buffer RWC	20 ml	80 ml
Buffer RW2*	20 ml	2 x 50 ml
RNase Free Water	10 ml	30 ml

Storage and Stability

MagZol Reagent should be stored at 2-8°C upon arrival. However, short-term storage (up to 24 weeks) at room temperature (15-25°C) does not affect its performance. The remaining kit components can be stored at room temperature (15-25°C) and are stable for at least 18 months under these conditions.

The Kit is designed for purification of total RNA, including miRNA and other small RNA molecules (18nt), from cultured cells and various animal and human tissues, including difficult-to-lyse tissues samples. Alternatively, a miRNA-enriched fraction and a total RNA (>200nt) fraction can be purified separately. This Kit combines phenol/guanidine-based lysis of samples and silica membrane–based purification of total RNA. MagZol Reagent, included in the kits, is a monophasic solution of phenol and guanidine thiocyanate, designed to facilitate lysis of tissues, to inhibit RNases, and also to remove most of the cellular DNA and proteins from the lysate by organic extraction.

Description

The ExcelTaq™ 5X PCR Master Dye Mix is a ready-to-use mixture for amplifying targeted DNA fragments. It is designed to serve as ready-to-use master mix for virtually all PCR applications. The mixture contains all components for PCR with the exception of template and primers. This not only saves valuable time in the laboratory, but also reduces the number of pipetting and reagent handling errors. The PCR Master Dye Mix is supplied as a 5X concentrated ready-to-use mix, that is a mixture of recombinant Taq DNA polymerase, reaction buffer, MgCl₂, dNTP, enzyme stabilizer and PCR-friendly loading dye solution containing tracking dye (Bromophenol blue) enabling efficient amplification of template in PCR and allows the user to prepare a PCR reagent – loading dye master mix conveniently.

Features

• 5’→3’ DNA polymerase activity
• No detectable 3’→5′ exonuclease (proofreading) activity
• Generates PCR products with 3′-dA overhangs
• High yield PCR 
• High reproducibility
• Reduced  pipetting errors
• Includes tracking dye for direct loading after PCR

Applications 

• Routine PCR
• Colony PCR
• High throughput PCR
• Amplification of DNA fragments up to 8 kb
• Generation of PCR products for TA cloning
• DNA labeling

Storage

4°C for 6 months
-20°C for 24 months

The ExcelTaq™ 5X PCR Master Dye Mix is a ready-to-use mixture for amplifying targeted DNA fragments. It is designed to serve as ready-to-use master mix for virtually all PCR applications. The mixture contains all components for PCR with the exception of template and primers. This not only saves valuable time in the laboratory, but also reduces the number of pipetting and reagent handling errors. The PCR Master Dye Mix is supplied as a 5X concentrated ready-to-use mix, that is a mixture of recombinant Taq DNA polymerase, reaction buffer, MgCl2, dNTP, enzyme stabilizer and PCR-friendly loading dye solution containing tracking dye (Bromophenol blue) enabling efficient amplification of template in PCR and allows the user to prepare a PCR reagent – loading dye master mix conveniently.