Molecular Biology Nucleic Acid Extraction Reagent for Rna
China Molecular Biology Nucleic Acid Extraction Reagent for Rna, Find details about China Nucleic Acid Extraction Solution, Rnaextract from Molecular Biology Nucleic Acid Extraction Reagent for Rna
Basic Info.
Model NO.
G3013-100ML
Habit Appellation
Special Reagent
Application
Industry, Scientific Research
Property
Biochemical Reagent
Trademark
Servicebio
Specification
100ML
Origin
China
Model NO.
G3013-100ML
Habit Appellation
Special Reagent
Application
Industry, Scientific Research
Property
Biochemical Reagent
Trademark
Servicebio
Specification
100ML
Origin
China
Product Desciption
Product introduction:
This product RNAextraction solution is a reagent used for total RNAextraction from cells or tissues. It uses guanidine isothiocyanate and other substances to rapidly lyse cells to release RNA and extract it into phnol. At the same time, it contains RNase inhibitors to prevent RNA degradation and maintain RNA integrity.
Package Contents:
Storage condition:
Wet ice bag transportation; Stored at 4ºC temperature, valid for 12 months.
Usage:
Self-prepared LF, isopropl alcohol, DEPC water, 75% ethanol (DEPC water preparation).
1. Lysis of a cell or tissue sample
Adhered cells: remove cell culture medium, clean cells with appropriate amount of pre-cooled PBS, and blot out PBS as much as possible. Add RNAextract to the well plate according to the amount of 1 mL RNAextract per 6-well plate and 0.5mL RNAextract per 12-well plate, shake gently to make the cells at the bottom of the plate fully contact the RNAextract, blow the cells with pipette gun to make the cells lyse, then transfer to the centrifuge tube, and let stand at room temperature for 5 min.
Suspend cells: Cells were collected by centrifugation, and the supernatant was removed as far as possible. 1mL RNAextract was added to every 1-10x106 cells, and the cells were blown several times with pipette gun to cleavage, then the cells were left standing at room temperature for 5min.
Tissue sample: cut the tissue into small pieces and put them into a pre-cooled homogenizer. Add 1mL RNAextract for every 100mg of tissue and homogenize until no tissue pieces are visible. The tissue fragments were separated by centrifugation at 12000g at 4ºC for 10min, and the supernatant was transferred to a new centrifuge tube.
2. RNAextraction: 380ul LF was added to each 1mL RNAextract, and then the centrifuge tube was shaken violently for 15s by eddy mixing or upside-down, and left for 3min at room temperature.
3. Centrifugation: centrifuge at 12000 g at 4ºC for 15 min, carefully transfer the upper colorless aqueous phase to the new centrifuge tube, and absorb 500-550ul per ml of extract.
4. Precipitated RNA: 550ul isoproanol was added to the aqueous phase extract tube collected above, mixed slightly upside down for several times, and precipitated at -20ºC for 15min.
5. Centrifugation: centrifuged at 4ºC for 10min at 12000g, white precipitation was visible at the bottom of the tube, which was total RNA, and supernatant was discarded.
6. Washing: Add 1 mL 75% ethanol and mix it upside down. After centrifugation at 12000g for 5min at 4ºC, the supernatant was discarded and then centrifuged briefly at a high speed (5000g for 3-5s). The liquid was carefully sucked out with a micro suction head.
7. RNA dissolution: The centrifuge tube with RNA precipitation was left open for 3-5min. After the RNA was slightly dried, 20ul DEPC water was added to make the RNA fully dissolved, which could be used for concentration detection and follow-up experiments. Be careful not to dry the RNA too much, otherwise it will be difficult to dissolve and will affect the A260/280 ratio.
Notes:
1. All pipette tips and centrifuge tubes shall be free of RNase contamination. You can purchase sterile enzyme-free consumables from our company.
2. This reagent contains guanidine isothiocyanate and resteamed phnol. Avoid contact with skin or inhalation.
3. Please wear lab clothes and disposable gloves.
This product RNAextraction solution is a reagent used for total RNAextraction from cells or tissues. It uses guanidine isothiocyanate and other substances to rapidly lyse cells to release RNA and extract it into phnol. At the same time, it contains RNase inhibitors to prevent RNA degradation and maintain RNA integrity.
Package Contents:
| Cat. No. | Product description | Volume |
| G3013 | RNAextraction solution | 100mL |
Storage condition:
Wet ice bag transportation; Stored at 4ºC temperature, valid for 12 months.
Usage:
Self-prepared LF, isopropl alcohol, DEPC water, 75% ethanol (DEPC water preparation).
1. Lysis of a cell or tissue sample
Adhered cells: remove cell culture medium, clean cells with appropriate amount of pre-cooled PBS, and blot out PBS as much as possible. Add RNAextract to the well plate according to the amount of 1 mL RNAextract per 6-well plate and 0.5mL RNAextract per 12-well plate, shake gently to make the cells at the bottom of the plate fully contact the RNAextract, blow the cells with pipette gun to make the cells lyse, then transfer to the centrifuge tube, and let stand at room temperature for 5 min.
Suspend cells: Cells were collected by centrifugation, and the supernatant was removed as far as possible. 1mL RNAextract was added to every 1-10x106 cells, and the cells were blown several times with pipette gun to cleavage, then the cells were left standing at room temperature for 5min.
Tissue sample: cut the tissue into small pieces and put them into a pre-cooled homogenizer. Add 1mL RNAextract for every 100mg of tissue and homogenize until no tissue pieces are visible. The tissue fragments were separated by centrifugation at 12000g at 4ºC for 10min, and the supernatant was transferred to a new centrifuge tube.
2. RNAextraction: 380ul LF was added to each 1mL RNAextract, and then the centrifuge tube was shaken violently for 15s by eddy mixing or upside-down, and left for 3min at room temperature.
3. Centrifugation: centrifuge at 12000 g at 4ºC for 15 min, carefully transfer the upper colorless aqueous phase to the new centrifuge tube, and absorb 500-550ul per ml of extract.
4. Precipitated RNA: 550ul isoproanol was added to the aqueous phase extract tube collected above, mixed slightly upside down for several times, and precipitated at -20ºC for 15min.
5. Centrifugation: centrifuged at 4ºC for 10min at 12000g, white precipitation was visible at the bottom of the tube, which was total RNA, and supernatant was discarded.
6. Washing: Add 1 mL 75% ethanol and mix it upside down. After centrifugation at 12000g for 5min at 4ºC, the supernatant was discarded and then centrifuged briefly at a high speed (5000g for 3-5s). The liquid was carefully sucked out with a micro suction head.
7. RNA dissolution: The centrifuge tube with RNA precipitation was left open for 3-5min. After the RNA was slightly dried, 20ul DEPC water was added to make the RNA fully dissolved, which could be used for concentration detection and follow-up experiments. Be careful not to dry the RNA too much, otherwise it will be difficult to dissolve and will affect the A260/280 ratio.
Notes:
1. All pipette tips and centrifuge tubes shall be free of RNase contamination. You can purchase sterile enzyme-free consumables from our company.
2. This reagent contains guanidine isothiocyanate and resteamed phnol. Avoid contact with skin or inhalation.
3. Please wear lab clothes and disposable gloves.
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