Strand Separation - Qiagen PyroMark Q96 ID User Manual

5. Seal the PCR plate using strip caps. Ensure that no
6. Agitate the PCR plate constantly for at least 5–10 min
7. Proceed with strand separation, see next section.
5.4.4

Strand separation

WARNING Hazardous chemicals
The Denaturation Solution used with the vacuum
workstation contains sodium hydroxide, which is irritating
to eyes and skin.
Always wear safety glasses, gloves, and a lab coat.
The responsible body (e.g., laboratory manager) must take
the necessary precautions to ensure that the surrounding
workplace is safe and that the instrument operators are not
exposed to hazardous levels of toxic substances (chemical
or biological) as defined in the applicable Safety Data
Sheets (SDSs) or OSHA,* ACGIH,
For more information, visit www.qiagen.com/safety.
Venting for fumes and disposal of wastes must be in
accordance with all national, state, and local health and
safety regulations and laws.
* OSHA: Occupational Safety and Health Administration (United States of America).
† ACGIH: American Conference of Government Industrial Hygienists (United States of America).
‡ COSHH: Control of Substances Hazardous to Health (United Kingdom).
PyroMark Q96 ID User Manual 01/2016
Note: If performing multiplex analysis of pooled PCR
products, use 5–10 µl of each product and reduce water
volume to maintain the total volume. The total volume
should be 80 µl after addition of the master mix and PCR
product.
leakage is possible between the wells.
using a mixer (1400 rpm).
Note: Sepharose beads sediment quickly and capturing
of beads must take place immediately once the agitation
is complete. If more than 1 minute has elapsed, agitate
the plate again before capturing the beads.
Note: During immobilization, prepare the Vacuum
Workstation for the sample preparation.
Operating Procedures
† or COSHH ‡ documents.
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