Blood RNA Isolation Kit
Kit Description:
Cat.No.RE-04011/04013
For total RNA purification from whole blood 104 ≤ White Blood Cells ≤ 107
Rapidly isolate and purify blood RNA from white blood cells.
-No need to worry about RNA degradation. The whole kit is RNase-Free
-Simple—all operations are completed at room temperature
-Fast—operation can be completed in 20 minutes
-High RNA yield: RNA-only Column and unique formula can efficiently purify RNA
-Safe—no organic reagent used
-Large sample processing capacity—up to 200μl samples can be processed each time.
-High quality—the purified RNA is highly pure, free of protein and other impurities, and can meet various downstream experimental applications.
Descriptions
The kit adopts the spin column and formula developed by our company, which can efficiently extract high-purity and high-quality total RNA from anticoagulated whole blood. The kit provides red blood cell lysate (Buffer RCL), which can quickly and efficiently lyse red blood cells and retain white blood cells. The efficient DNA-Cleaning Colunm can easily separate the supernatant and cell lysates and adsorb and remove genomic DNA. The operation is simple and time-saving ; The RNA-only Column can efficiently bind RNA, and with a unique formula, it can process a large number of samples at the same time.
The whole system RNase-Free makes the extracted RNA non-degradable; Buffer RW1 and Buffer RW2 buffer washing system makes the obtained RNA free of protein, DNA, ions and organic compound pollution.
Kit Contents
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Blood Total RNA Isolation Kit |
||
|
Kit composition |
RE-04011 |
RE-04013 |
|
50 times |
200 times |
|
|
Buffer RCL (10×) |
52.5 mL |
210mL |
|
Buffer BRL1* |
30mL |
120mL |
|
Buffer BRL2 |
18mL |
66mL |
|
Buffer RW1* |
25mL |
100mL |
|
Buffer RW2 |
24mL |
96mL |
|
RNase-Free ddH 2 O |
10mL |
40mL |
|
RNA-only Column |
50 sets |
200 sets |
|
DNA-Cleaning Column |
50 sets |
200 sets |
|
manual |
1 copy |
1 copy |
Features&advantages
-No need to worry about RNA degradation. The whole kit is RNase-Free.
-Simple—all operations are completed at room temperature.
-Fast—operation can be completed in 20 minutes.
-High RNA yield: RNA-only Column and unique formula can efficiently purify RNA.
-Safe—no organic reagent used.
-Large sample processing capacity—up to 200μl samples can be processed each time.
-High quality—the purified RNA is highly pure, free of protein and other impurities, and can meet various downstream experimental applications.
Kit parameters
Kit application:
It is suitable for the extraction and purification of total RNA from mammalian whole blood.
Work flow
Storage conditions
Buffer RCL (10×) should be stored at 2-8 ℃ ; other components of the kit can be stored at room temperature (15-25 ℃ ) under dry conditions, and can be stored for 12 months. Buffer BRL1 can be stored at 4 ℃ for 1 month after adding β-mercaptoethanol (optional) .
Note: If stored at low temperature, the solution is prone to precipitation. Be sure to place the solution in the kit at room temperature for a period of time before use. If necessary, preheat it in a 37° C water bath for 10 minutes to dissolve the precipitate, and mix well before use.
Guides for problems analysis
The following is an analysis of the problems that might be encountered in the extraction of viral RNA. We wish it would be helpful to your experiment. In addition, for other experimental or technical problems other than operating instructions and problem analysis, we have dedicated technical support to help you. Contact us if you need at : 028-83361257or E-mail：Tech@foregene.com。
No RNA can be extracted or the yield of nucleic acid is low
There are usually many factors that affect recovery efficiency, such as: sample RNA content, method of operation, elution volume, etc.。
Analysis of common causes：
1.Ice bath or low-temperature (4 ° C) centrifugation during operation.
Suggestion: Room temperature (15-25 ° C) operation, never ice bath and low temperature centrifuge.
2. Improper sample storage or sample storage for too long.
Suggestion: Store samples at -80 ° C or freeze in liquid nitrogen, and avoid repeated freeze-thaw use; try to use freshly collected samples for RNA extraction.
3.Insufficient sample lysis
Recommendation: Please ensure that the sample and the working solution (Linear Acrylamide) have been thoroughly mixed and incubated for 10 min at room temperature (15-25 ° C)
4.The eluent was added incorrectly
Recommendation: Make sure that RNase-Free ddH2O is added to the middle of the membrane of the purification column
5.Improper volume of anhydrous ethanol in Buffer viRW2
Suggestion: Please follow the instructions, add the correct volume of anhydrous ethanol to Buffer viRW2 and mix them well before using the kit。
6.Improper sample usage.
Suggestion: 200µl of sample per 500μl of Buffer viRL. Excessive sample volume will result in reduced RNA extraction rate.
7.Improper elution volume or incomplete elution.
Suggestion: The eluent volume of the purification column is 30-50μl; if the elution effect is not satisfactory, it is recommended to add pre-heated RNase-Free ddH2O and extend the time placing at room temperature, such as 5-10min
8.Purification column has ethanol residue after rinsing in Buffer viRW2.
Suggestion: If ethanol still remains after rinsing in Buffer viRW2 and empty-tube centrifugation for 2min, the purification column can be left at room temperature for 5min after empty-tube centrifugation to fully remove remaining ethanol.
The degradation of purified RNA molecules
The quality of the purified RNA is related to factors such as sample storage, RNase contamination, and operation.
Analysis of common causes：
1.The collected samples were not saved in time.
Suggestion: If the sample is not used in time after collection, please store it at -80 ℃ or liquid nitrogen immediately. For the extraction of RNA molecules, try to use freshly collected samples whenever possible.
2.Collected samples were freezing and thawing repeatedly.
Suggestion: Avoid repeated freezing and thawing (no more than once) during sample collection and storage, otherwise the yield of nucleic acid will decrease.
3.RNase was introduced in the operating room or no disposable gloves, masks, etc. were worn.
Suggestion: The extraction of RNA molecules experiment is best performed in a separate RNA operation room, and the experimental table is cleaned before the experiment. Wear disposable gloves and masks during the experiment to avoid RNA degradation caused by RNase introduction.
4.The reagent is contaminated by RNase during the use.
Suggestion: Replace with new Viral RNA Isolation Kit for related experiments.
5.The RNase contamination of the centrifuge tubes, pipette tips, etc. Suggestion: Make sure that the centrifuge tubes, pipette tips, and pipettes are all RNase-Free.
The purified RNA molecules affected downstream experiments
The RNA molecules purified by the purification column will affect downstream experiments if there are too much salt ions or proteins, such as: reverse transcription, Northern Blot, etc.。
1.There are remaining salt ions in the eluted RNA molecules.
Recommendation: Make sure that the correct volume of anhydrous ethanol has been added to Buffer viRW2, and wash the purification column twice according to the correct centrifugation speed on the operating instructions；If there are still salt ions remaining, you can add Buffer viRW2 to the purification column, and leave it at room temperature for 5min. Then perform centrifugation to remove salt ions contamination to the greatest extent
2.There are remaining ethanol in the eluted RNA molecules
Suggestion: once confirming that purification columns have been rinsed by Buffer viRW2, perform empty-tube centrifugation according to the centrifugal speed on the operating instructions. If there is still ethanol remaining, it can be left for 5 minutes at room temperature after an empty-tube centrifugation to remove the remaining ethanol to the greatest extent.
Instruction Manuals:
Viral RNA Isolation Kit Instruction Manual
