N-(t-butyl ester-PEG4)-N-bis(PEG4-Propargyl) is a trifunctional PEG linker that combines a t-butyl ester with two terminal alkynes. The alkynes can be applied in copper-click chemistry with azides to form stable triazole linkages with a target molecule, while the t-butyl ester can be deprotected and reacted with amines or alcohols to form amides or esters.

N-(t-butyl ester-PEG4)-N-bis(PEG4-Propargyl) is a trifunctional PEG linker that combines a t-butyl ester with two terminal alkynes. The alkynes can be applied in copper-click chemistry with azides to form stable triazole linkages with a target molecule, while the t-butyl ester can be deprotected and reacted with amines or alcohols to form amides or esters.

Introduction

HiPure Soil RNA Kit is suitable for extracting high-purity microbial total RNA from soil samples. The kit adopts silica gel column purification technology and original humic acid adsorbent technology. It is suitable for extracting high-yield and high-purity total RNA from various soil samples, such as forest soil, grassland soil, mining soil, sediment and so on. The obtained RNA can be directly used in RT-PCR, Northern blot, poly-A + purification, nucleic acid protection and in vitro translation.

Details

Specifications

Features	Specifications
Main Functions	Isolation total RNA from 500 mg soil sample
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	Forest soil, grassland soil, mining area soil, sediment and other samples
Sample amount	500 mg
Elution volume	≥30μl
Time per run	≤60 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 as guanidine hydrochloride or guanidine isothiocyanate), the filter membrane can adsorb nucleic acid through hydrogen bond and electrostatic, while protein and other 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.

Advantages

• High quality – high purity total RNA can be directly used in various sensitive downstream applications
• Fast – extraction of several samples can be completed in 60 minutes by column purification method
• Safe – no phenol chloroform extraction required
• Sensitive – RNA can be purified at the level of PG

Kit Contents

Contents	R418302	R418303
Purification Times	50 Preps	250 Preps
HiPure RNA Micro Columns	50	250
2ml Collection Tubes	100	500
gDNA Filter Column	50	250
2ml Beads Tubes	50	250
Buffer SOL	30 ml	150 ml
Buffer SDS	4 ml	15 ml
Buffer PHC	30 ml	150 ml
Buffer GDP	40 ml	150 ml
Buffer RW1	50 ml	200 ml
Buffer RW2 *	20 ml	2 x 50 ml
RNase Free Water	10 ml	30 ml

Storage and Stability

Buffer PHC should be stored at 2–8°C upon arrival. However, short-term storage (up to 12 weeks) at room temperature (15–25°C) does not affect their performance. The remaining kit components can be stored dry at room temperature (15–25°C) and are stable for at least 18 months under theseconditions.The entire kit can be stored at 2–8°C, but in this case buffers should be redissolvedbefore use. Make sure that all buffers are at room temperature when used.

Experiment Data

HiPure Soil RNA Kit is suitable for extracting high-purity microbial total RNA from soil samples. The kit adopts silica gel column purification technology and original humic acid adsorbent technology. It is suitable for extracting high-yield and high-purity total RNA from various soil samples, such as forest soil, grassland soil, mining soil, sediment and so on. The obtained RNA can be directly used in RT-PCR, Northern blot, poly-A + purification, nucleic acid protection and in vitro translation.

Overview

• Quantified standard to be used as a positive control or PCR quantification standard
• Vigorously quantified using multiple methods
• Includes partial sequence of STX-1 subunit A, and STX-2 subunit A genes (please refer to product insert for more details)
• Compatible with E. coli O157:H7 TaqMan PCR Detection Kit and E. coli O157:H7 TaqMan PCR Detection Probe/Primer and Control Set

E. coli O157:H7 is a rod-shaped, gram negative bacterium. It is an enterohemorrhagic strain of the common E. coli bacterium and infection by the O157:H7 strain is commonly associated with hemorrhagic colitis. E. coli O157:H7 is recognized by its somatic (cell wall) antigen (O157) and its flagella antigen (H7). In addition, E. coli O157:H7 is known to produce Shiga-like toxins, which cause severe symptoms. While most patients can recover from the infection, up to 15% of the patients may develop hemolytic uremic syndrome, a type of kidney failure that could be fatal. Infection of E. coli O157:H7 usually results from consumption of poorly prepared food including undercooked meat (particularly ground beef), untreated water or raw unpasteurized milk.

Norgen’s E. coli O157:H7 Quantified Bacterial DNA Standards are cloned fragments of the two Shiga-like toxin regions purified using Norgen’s sample preparation technology. They include partial sequences of STX-1 subunit A gene and STX-2 subunit A gene. Please refer to Appendix A for sequence information. The purified DNA is quantified vigorously using multiple methods including spectrophotometry, gel densitometry and real-time PCR. It is intended to be used as positive controls or PCR quantification standards for E. coli O157:H7. This product is compatible with Norgen Biotek’s E. coli O157:H7 Taqman PCR Detection Kit (Cat. TM41350) and E. coli O157:H7 Taqman PCR Detection Probe/Primer and Control Set (Cat. TM41310).

Details

Supporting Data

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Volume Provided	250 μL (for STX1)250 μL (for STX2)
DNA Quantity	2 x 104 copies per μL

Storage Conditions
Upon receipt, store Norgen’s E. coli O157:H7 Quantified Bacterial DNA Standards at -20°C or lower. Avoid multiple freeze-thaw cycles. If needed, prepare smaller working aliquots and store at -20°C or lower.