RNA Extraction
α+ Solution™ Blood / Cultured Cell Total RNA Mini Kit
Catalog #: APBRK004 / APBRK050 / APBRK100
Manufacturer: ALPHAGEN
Price: on inquiry
- Principle
- mini spin column (silica matrix)
- Sample size
- 300 μl
- Operation time
- 30 ~ 60 minutes
- Storage
- Store at room temperature (15~ 25 ℃) for 2 year.
Ordering information
| APBRK004 | 4 preps (sample) |
|---|---|
| APBRK050 | 50 preps |
| APBRK100 | 100 preps |
Storage: Store at room temperature (15~ 25 ℃) for 2 year.
Component
| APBRK 004 (4 preps) |
APBRK 050 (50 preps) |
APBRK 100 (100 preps) |
|
| RL Buffer | 15 ml x 2 | 120 ml | 240 ml |
| RB Buffer | 1.5 ml x 2 | 25 ml | 45 ml |
| Wash Buffer 1 | 1.5 ml x 2 | 30 ml | 60 ml |
| Wash Buffer 2 * (concentrate) | 1.5 ml | 15 ml | 35 ml |
| RNase-free Water | 0.5 ml | 6 ml | 6 ml |
| Filter Column | 4 pcs | 50 pcs | 100 pcs |
| RB Mini Column | 4 pcs | 50 pcs | 100 pcs |
| Collection Tube | 8 pcs | 100 pcs | 200 pcs |
| Elution Tube | 4 pcs | 50 pcs | 100 pcs |
| User Manual | 1 | 1 | 1 |
Brief Prodecure
Specification
※ Principle : mini spin column (silica matrix)
※ Sample size : 300 μl
※ Operation time : 30 ~ 60 minutes
※ Binding capacity : up to 100 μg total RNA / column
※ Expected yield : 1 μg
※ Column applicability : centrifugation and vaccum
※ Minimum elution volume : 40 μl
Important Notes :
※ Make sure everything is RNase-free when handling RNA.
※ Buffers provided in this system contain irritants. Wear gloves and lab coat when handling these buffers.
※ Caution: ß-mercaptoethanol is hazardous to human health. perform the procedures involving RB Buffer or PRB Buffer in a chemical fume hood.
※ Add required volume of RNase-free ethanol (96~100%) to Wash Buffer 2 when first use.
※ Dilute RNase-free DNase 1 in reaction buffer (1M NaCl, 10 mM MnCl2, 20 mM Tris-HCl, pH 7.0 at 25ºC) to final conc. = 0.5 U/μl.
Trouble shooting
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Problems |
Possible reasons |
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Low yield |
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Carryover of erythrocytes. |
Incomplete removal of the supernatant will interfere with lysis and subsequent binding of RNA to the RB column, resulting in lower yield. |
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Too much starting sample used may cause insufficient lysis and homogenization. |
In Step.1 sample preparation, reduce the amount of starting sample or increase the volume of RL Buffer. |
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Incorrect RNA elution step. |
Ensure that Elution Buffer was added and absorbed to the center of the PD Column matrix. |
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Ethanol carryover |
Ensure that centrifuge at ≥ 8000 x g (≥10,000 rpm) for 3 min to dry the RB column membrane in Step.6 Dry Column. |
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Incorrect preparation of Wash 2 Buffer |
Ensure that the correct volume of ethanol (96 ~ 100 %) was added to Wash 2 Buffer before use. |
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RNA degraded |
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Age of blood. |
Please use the fresh blood sample. |
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Residual DNA |
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DNA Contamination. |
Please follow the Optional step to remove DNA. |
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RNA does not perform well in downstream experiments |
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Salt carryover during elution. |
Ensure that Wash 2 Buffer is at room temperature (15–25°C). |
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Ethanol carryover. |
Perform the step. 6 in the protocol (centrifuge at ≥ 8000 x g or ≥ 10,000 rpm) to remove all traces of ethanol before eluting. |
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Documents
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