It is suitable for the extraction of RNA from blood, plasma, serum, lymph and other samples. The silica-based material used in the centrifugal adsorption column is a unique new material of our company, which can maximize the recovery of high-purity RNA with our unique lysis solution formula. The extracted RNA is of high purity, stable and reliable quality, and can be applied to various downstream application experiments, such as RT-PCR, RT-qPCR, in vitro transcription, molecular cloning, etc.
How to operate
1. Sample preprocessing:
Transfer 250 μL of blood (or serum, plasma, cerebrospinal fluid, etc.) to a 1.5 mL RNase-free centrifuge tube.
【Note】: When it is less than 250 μL, it needs to be supplemented with PBS or normal saline.
Add 750 μL of Lysate LB, pipetting repeatedly, and shake vigorously to mix.
Let stand at room temperature for 5 min.
Add 200 μL of chloroform (provided by yourself) and mix vigorously for 15 sec.
Let stand at room temperature for 2 min.
Centrifuge at 12,000 rpm for 10 min at 4°C to separate the samples. Transfer the upper aqueous phase to a 1.5 mL RNase-free centrifuge tube.
【Note】: The sample will be divided into three layers, the lower layer is the organic phase, the middle layer and the upper layer are colorless aqueous phase, and RNA exists in the upper aqueous phase.
【Note】: The upper layer volume is about 70% of the total amount of lysate LB added. If 750 μL of Lysis LB is added, the upper aqueous phase is about 525 μL. It is recommended to pipette 500 μL to prevent DNA contamination by pipetting into the middle layer.
Add 0.5 times the volume of absolute ethanol (provided by yourself) and mix by inversion.
【Note】: Precipitation is a normal phenomenon.
2. RNA extraction:
Set the RNA adsorption column B3 into a 2 mL collection tube and set aside.
Add the above pretreatment mixture to RNA adsorption column B3, centrifuge at 12,000 rpm for 1 min, and discard the waste solution.
Add 500 μL of deproteinized solution PL, centrifuge at 12,000 rpm for 30 sec, and discard the waste solution.
Put the RNA adsorption column B3 back into the collection tube, add 500 μL of washing solution W*, centrifuge at 12,000 rpm for 30 sec at room temperature, and discard the waste solution.
【Note】: Make sure that the rinse solution W* has been added with absolute ethanol.
Repeat step 4 again.
Put the RNA adsorption column B3 back into the collection tube and centrifuge the empty column at 12,000 rpm for 2 min at room temperature to remove residual wash solution W*.
Put RNA adsorption column B3 into a new 1.5 mL RNase-free centrifuge tube, add 30-50 μL RNase-free H2O to the center of RNA adsorption column B3, and leave at room temperature for 2 min. Then centrifuge at 12,000 rpm for 1 min. Collect the filtrate, which is the RNA solution.
【Note】: The recovery yield can be improved by the following methods: ①Preheat RNase-free H2O at 65℃; ②Put the RNA filtrate on the column again, place it at room temperature for 2 minutes, and then elute.
RNA solution can be stored at -80°C for a long time.
Warm tip: the products supplied by Beijing Beike Xincai Technology Co., Ltd. are only used for scientific research, not for human body |
Item ID |
CAS |
ID |
Pack |
Parameter |
Stock |
Make up |
Price |
BK2022031705 |
|
BK2022031705 |
6X16T |
|
100 |
|
$480 |