Description

Buffer PLY, Wash Buffer, DEPC-Treated ddH2O, ezBind Columns, Collection Tubes, User Menu

Introduction

The EZgene^TM Viral DNA/RNA kit provides an easy and reliable method for isolating total viral DNA/RNA from plasma, serum, whole blood, urine and cell culture supernatant. This procedure has been tested for isolating nucleic acids from Hepatitis A, Hepatitis C and HIV. The isolated DNA/RNA can be used for PCR, RT-PCR and other downstream applications.

Storage and Stability

All other components can be stored at room temperature. All kit components are guaranteed for 1 year from the date of purchasing.

 

Kit Contents

Catalog#	VR6511-00	VR6511-01	VR6511-02
Preps	4	50	250
Buffer PLY	2.4 mL	28 mL	130 mL
Wash Buffer *	2 mL	15 mL	3 x 24 mL
DEPC-Treated ddH2O	500 µL	10 mL	50 mL
ezBind Columns	4	50	250
Collection Tubes	8	100	500
User Menu	1	1	1

*Add 8 mL (VR6511-00) or 60 mL (VR6511-01) or 96  mL (VR6511-02) 100% ethanol into each Wash Buffer bottle before use.

Before Starting

Prepare all components and get all necessary materials ready by examining this instruction booklet and become familiar with each step.

Important

•  Add 1% volume of β-mercaptoethanol to Buffer PLY before use and store at 4 ^oC.
•  Add 8 mL (VR6511-00) or 60 mL (VR6511-01) or 96 mL (VR6511-02) 100% ethanol into each Wash Buffer bottle before use.

Materials supplied by users

•  Tabletop microcentrifuge
•  1.5 mL sterile tubes
•  100% ethanol

Note: Perform all steps including centrifugation at room temperature

Trouble Shooting Guide

Problem	Possible reason	Suggested Improvement
Low A260/A280 ratios	Protein contamination	Do a Phenol:Chloroform extraction. Loss of total RNA (up to 40%) should be expected.
Low A260/A280 ratios	Guanidine Thiocyanate contamination	Add 2.5 volumes of ethanol and 0.1M NaCl (final concentration) to precipitate RNA. Incubate for 30 min at -20°C. Centrifuge at 13,000 rpm for 15 min at 4°C. Resuspend the RNA pellet in DEPC-treated water.
Low Yield	RNA in sample degraded	Freeze samples immediately in liquid nitrogen and store at -70°C after collect it.
Low Yield	The binding capacity of the membrane in the spin column was exceeded	Use of too much sample exceeding the binding capacity of spin column will cause the decreasing of total RNA yield.
Low Yield	Ethanol not added to buffer	Add ethanol to the Wash Buffer.
Genomic DNA contamination	Too much total RNA sample was used in RT-PCR.	Reduce total RNA amount used in RT-PCR to 50-100 ng.