商品介紹
BDCK011 Tissue Total RNA Mini Kit
簡易Protocol
※ Principle : mini spin column (silica matrix)
※ Sample size : Please see Table. Sample amount and Yield.
※ Operation time : 30 ~ 60 minutes
※ Binding capacity : up to 100 μg total RNA/ column
※ Expected yield : Please see Table. Sample amount and Yield.
※ 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.
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Problems |
Possible reasons |
Solutions |
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Low 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 RB Buffer. |
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Incorrect RNA elution step. |
Ensure that Elution Buffer was added and absorbed to the center of the RB 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 Sample. |
Please use the fresh 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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