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Purification
The finished library is purified from PCR components that can interfere with quantification.
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dd 160 µl of column binding buffer (CBS) to the reaction, mix well, and transfer the
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solution to a column placed in a 2 ml collection tube. Centrifuge for 1 minute at 12,000
x g at 18 °C.
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Apply 200 µl of column wash buffer (CW) to the column and centrifuge for 1 minute.
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Repeat this washing step once (for a total of two washes).
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emove the column and transfer to a fresh collection tube. Centrifuge for 2 minutes at
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12,000 x g at 18 °C to dry the column.
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Transfer the column to a new 1.5 ml tube and apply 15 µl elution buffer (EB ) to the
column. Incubate at room temperature for 1 minute and centrifuge for 2 minutes at
12,000 x g at 18 °C to elute the library.
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t this point, the libraries are finished and ready for quality control (Appendix C, p.22),
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pooling (for multiplexed SENSE libraries; see Appendix D, p.24), and cluster generation.
LEXOGEN · mRNA-Seq Library Prep Kit · User Guide