Aquatic Animal Virgin Genomic DNA/RNA Extraction Kit Instruction Manual

Aquatic animal virus genomic DNA/RNA extraction kit

First, the introduction

The aquatic animal viral genomic DNA/RNA extraction kit is suitable for rapidly extracting high purity viral genomic nucleic acids from aquatic animal tissue homogenates. The kit is based on silica gel column purification technology. It does not require the use of toxic phenol chloroform extraction or time-consuming alcohol precipitation. The obtained nucleic acid can be directly used in PCR/RT-PCR, Sorter hybridization/Northern hybridization, and Downstream experiments in series such as LAMP/RT-LAMP.

• principle

The silica gel column used in this kit is based on a high-binding glass fiber filter. The filter membrane can adsorb nucleic acids by physicochemical interactions such as hydrogen bonding and static electricity under the condition of high concentration ionizing agent (such as guanidine hydrochloride or guanidinium isothiocyanate), while proteins and other impurities are not adsorbed and removed. The nucleic acid-adsorbing filter is washed to remove residual proteins and salts, and finally the DEPC can be used to treat the nucleic acid adsorbed on the water-eluting filter. The obtained nucleic acid has high purity and can be directly used in various downstream experiments.

The aquatic animal virus genomic DNA/RNA extraction kit is based on a silica gel column purification method. The sample is cleaved in the lysate and the nucleic acid is released into the lysate. The nucleic acid is adsorbed on the membrane of the silica gel column through a silica gel column in a high salt environment, and the protein is not adsorbed and removed. The salt is then washed by washing to remove the salt, and finally the nucleic acid is eluted by the nuclease-free eluate.

Third, the composition

Note : The washing solution was added to 40 ml of absolute ethanol before the initial use and stored at room temperature.

Fourth, the shelf life

The aquatic animal viral genomic DNA/RNA extraction kit component can be stored dry for 12 months at room temperature (15-25 ° C). It should be placed at 2-8 °C for long-term storage.

V. Materials and tools that need to be prepared

• Sterile saline
• Absolute ethanol
• Clean scorpion and scissors
• Homogenizer, homogenized bag or disposable grinding rod
• Micropipette (100-1000μl, 10-100μl)
• Sterilized centrifuge tubes without DNase and RNase (1.5ml or 2ml)
• Sterilized Tip Head without DNase and RNase
• Vortex oscillator
• Centrifuge (speed ≥ 10,000 rpm)

Method for rapidly extracting DNA/RNA from aquatic animal virus

The protocol uses centrifugation and is suitable for rapid extraction of viral genomic DNA/RNA from aquatic animal tissue samples. The following steps are all performed at room temperature.

• Take appropriate amount of fresh sample tissue to be tested and add 1~5 times the volume of normal saline for homogenization. Take at least 500 μl of the homogenate into a 1.5 ml centrifuge tube.
• The host cell tissue nucleic acid was removed by centrifugation at 10,000 rpm for 5 min.
• 100 μl of the supernatant was taken to a new centrifuge tube, 300 μl of the lysate was added to the supernatant, mixed by inversion, and allowed to stand for 5-10 min to lyse the virus.
• Add 200 μl of absolute ethanol to the lysate and vortex for 20 seconds.
• Tube A was placed in 2 ml tube B, and the lysate was all transferred to tube A and centrifuged at 10,000 rpm for 1 min.
• The filtrate in tube B was discarded, tube A was returned to tube B, 400 μl of washing solution was added to tube A, and centrifuged at 10,000 rpm for 1 min.
• Repeat step 6.
• The filtrate in tube B was discarded, tube A was returned to tube B, and centrifuged at 10,000 rpm for 3 min.
• Tube A was placed in a new 1.5 ml centrifuge tube.

• 30-50 μl of eluate was added to tube A, allowed to stand for 1-2 min, and centrifuged at 10,000 rpm for 1 min. The resulting solution was a viral genomic nucleic acid solution. If not used temporarily, store at -80 °C.

Seven, frequently asked questions

This list may help you solve the problems you encountered during the extraction process. If you have questions or have any suggestions for the kit, or if you have problems with molecular biology experiments, please contact us. We will do our best to help you solve problems.

If the above solution still does not solve your problem, please contact us.

YT-T15

YT-T15

YT-T15

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