Virus DNA/RNA Extraction Kit

 

Product Name	Cat.No.	Spec.
Virus DNA/RNA Extraction Kit	G3647-50T	50T

 

 

The kit is designed for rapid isolation of various viral DNA/RNA from serum, plasma, urine, supernatant of cell culture medium, virus stock and infected tissues. By making use of the characteristics of the nucleic acid adsorption column reversibly binding to the nucleic acid, coupled with a special buffer system, the nucleic acid is allowed to specifically bind to the membrane matrix,andproteins, cell fragments and other pollutants are effectively rinsed, and finally the nucleic acid is eluted from the adsorption column with Nuclease-free Water. The obtained high quality DNA/RNA can be directly used in downstream PCR, cDNA synthesis, RT-qPCR and other molecular biology experiments.

 

 

Proteinase K and Carrier RNA shipped with wet ice and stored at -20°C. Other reagents were shipped and stored at room temperature; valid for up to 1 year.

 

 

Component Number	Component	G3647-50T
G3647-1	BufferVLB	10 mL
G3647-2	Proteinase K	1 mL
G3647-3	Carrier RNA	150 μL
G3647-4	BufferVWA	12 mL
G3647-5	Buffer VWB	15 mL
G3647-6	Nuclease-free Water	12 mL
G3647-7	Spin Columns with Collection tubes	50
Manual		One copy

 

 

1. If BufferVLB and BufferVWA precipitates, please heat it at 37°C and use it when it is restored to room temperature.

2. Please add 18 mL anhydrous ethanol to BufferVWA and 60 mL anhydrous ethanol to BufferVWB before first use.

3. Carrier RNA can be sub-packed and stored at -20°C before first use, and repeated freezing and thawing should be avoided.

4. Please pre-cool the grinder if grinding tissue with a grinder.

 

 

1. Samples preparation:

a. Lysis of serum, plasma, urine, supernatant of cell culture medium and virus stock: Sampling 10-200 μL of serum, plasma, urine, supernatant of cell culture or virus stock. The initial amount of less than 200 μL can be replenished to 200 μL with PBS or Nuclease-free Water.

b. Lysis of infected tissue: Place 10~20 mg fresh or cryopreserved infected tissues in a 1.5 mL Nuclease-free centrifuge tube containing 2~3 pcs 3 mm grinding beads (recommended G0203) or use special grinding tubes (recommended HT-200-M). Immediately place the centrifuge tube or grinding tube containing the tissue in liquid nitrogen, and then grind the tissue thoroughly to homogenate at low temperature using a tissue grinder (recommended KZ-5F-3D, if the tissue is not thoroughly homogenized, the yield and quality of DNA/RNA will be affected). Add 200 μL PBS or Nuclease-free Water after grinding.

2. Add 200 μL BufferVLB,20 μL Proteinase K and 3 μL Carrier RNA. Aftermix thoroughly, incubate at 56°C for 10 min.

3. Add 200 μL anhydrous ethanol, mix upside down.

4. Transfer the solution from the previous step to Spin Column, centrifuge at 12,000 rpm for 1 min, and discard the filtrate from Collection tube.

5. Add 500 μL Buffer VWA to Spin Column, centrifuge at 12,000 rpm for 1 min, and discard the filtrate from Collection tube.

6. Add 700 μL Buffer VWB to the Spin Column (please add Buffer VWB along the Spin Column pipe wall to help flush the residual salt on the pipe wall). centrifuge at 12,000 rpm for 30 s, and discard the filtrate.

7. Repeat step6.

8. Put Spin Column on Collection tube, centrifuge at 12,000 rpm for 2 min.

9. Transfer Spin Column to a new Nuclease-free 1.5 mL centrifuge tube, open the lid for 3~5 min at room temperature, so that the residual ethanol of Spin Column can be evaporate completely.

10. Add30~50 μL Nuclease-free Water to the centrifugal tube,stand at room temperature for5min, centrifuge at 12,000 rpm for 2 min at room temperature to collect RNA solution. To get a higher concentration of RNA, you can also add the first eluent back to RNA Spin Column, then stand at room temperature for 5 min and centrifuge for 2 min to collect RNA again.

 

 

1. Please read the Product Manual carefully before use.

2. Due to the addition of Carrier RNA in the extraction process, the quantity can not be determined by electrophoresis or absorptometer.

3. The Carrier RNA provided by this kit is the RNA from E. coli, which is mainly to improve the recovery efficiency of trace nucleic acid and prevent the degradation of trace RNA. If the PCR primers have high homology with Carrier RNA, the primers can be redesigned.

4. Before elution, ethanol should be completely volatilized to avoid the influence of residual ethanol on downstream experiments.

5. Do not dry for a long time, so as not to affect the efficiency of nucleic acid elution.

6. For your safety and health, please wear a lab coat and disposable gloves.

 

For Research Use Only!