Cycle-Pure Spin Protocol Ur Supplied Equipment:
•Microcentrifuge capable of at least 13,000xg英语专业八级考试
•Nuclea-free 1.5 microcentrifuge tubes
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•Absolute ethanol (~96-100%)
•Optional: Sterile deionized water (or TE Buffer)gopast
boutiqueThings to do before starting:
•Prepare DNA Wash Buffer according to directions on page 4.
1. Perform agaro gel/ethidium bromide electrophoresis to analyze PCR product.大学英语六级词汇
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2.Determine the volume of the PCR reaction. Transfer the sample into a clean 1.5ml microcentrifuge tube and add 4-5 volumes of Buffer CP. For PCR products smaller than 200bp, add 6 volumes of Buffer CP.
Note: V olumes refers to the size of your PCR reaction. For example, if your PCR reaction is 100 µl an
d is smaller than 200bp, you would u 600 µl of Buffer CP.
honesty3. V ortex thoroughly to mix. Briefly spin the tube to collect any drops from the inside of the lid.
4. Place a HiBind DNA Mini Column into a provided 2 ml collection tube.
5. Add the mixed sample from step 3 to the HiBind DNA Mini Column and centrifuge at 13,000 x
g for 1 minute at room temperature. Discard the flow-through liquid and place the HiBind DNA Mini Column back into the same collection tube.
6. Add 700µl of DNA Wash Buffer and centrifuge at 13,000 x g for 1 minute. Discard the flow-through liquid and place the HiBind DNA Mini Column back into the same collection tube.
IMPORTANT: DNA Wash Buffer must be diluted with absolute ethanol before u. Refer to label for instructions. If refrigerated, DNA Wash Buffer must be brought to room temperature before u.
7. Add 500µl of DNA Wash Buffer and centrifuge at 13,000 x g for 1 minute. Discard the flow-through liquid and place the HiBind DNA Mini Column back into the same collection tube.
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8. Centrifuge the empty HiBind DNA Mini column for 2 min at maximal speed ( ≥13,000 x g ) to dry the column matrix.
Note: Do not skip this step, it is critical for the removal of ethanol from the HiBind DNA column.
妇女节英文9. Place the HiBind DNA Mini column into a clean 1.5ml microcentrifuge tube. Depending on the desire concentration of the final product, add 30-50 µl of Elution Buffer (10mM Tris, pH8.5) or water directly onto the center of column matrix.
