RNA extraction Step by action for RT-PCR|PCR Techniques part-1

We have actually begun a series on PCR methods
Today is our very first video on rna extraction action by action for covid-19(RT-PCR)

Spina RNA Viral Extraction Kit

Test Principle

This item is based upon silica gel filtration. The sample is lysed and absorbed with lysate and protease, RNA is launched into the lysate. Transfer to an adsorption plate and filter column. RNA is adsorbed on the membrane, while protein is not adsorbed and is gotten rid of with filtering. After cleaning proteins and other pollutants, RNA was lastly eluted with low-salt buffer.

Treatment Step by action
1. In a 1.5 ml centrifuge tube, include 400 ml Buffer VLF, 10 ul Proteinase K and 1ul Carrier RNA.
2. Transfer 200 ul of sample and mix by vortexing for 15 seconds. Sit at space temperature level for 5 minutes.
3. Get a brand-new Viral Mini column and move the sample to the column. Centrifuge at 10,000 rpm for 30-60 seconds
4. Dispose of the filtrate and location the column back into the collection tube. Include 500 ul Buffer CW to the column. Centrifuge at 10,000 rpm for 30-60 seconds.
5. Dispose of the filtrate and location the column back into the collection tube. Include 500 ul Buffer CW to the column. Centrifuge at 10,000 rpm for 30-60 seconds.
6. Dispose of the filtrate and location the column back into the collection tube. Centrifuge the column at 13,000 rpm for 3 minutes to dry the column.
7. Transfer the column to a brand-new 1.5 ml centrifuge tube.
8. Include 30-50 ul Buffer AVE to the center of the membrane of the column. Centrifuge at 13,000 rpm for 1 minute
9. Dispose of the column and shop the RNA at -80 ° C.
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