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PippinHT Operations Manual
2-1
460005 Rev G
2
Keys to Successful Collections
It is recommended that users closely follow the instructions outlined in this manual until
they become thoroughly familiar with the system. The Quick Guide (included with
cassettes) provides protocol reminders, but is not intended for beginning users.
Keys to successful collections are outlined in the following sections:
•
Purity of the input sample
(Section 6)
•
Optical Calibration
(Section 8)
•
Preparation of the cassettes
(Section 9)
•
Sample loading
(Section 11)
In addition, extra steps should be taken to improve recovery of high molecular weight
collections using High-Pass protocols with 0.75% agarose cassettes. These
recommendations are outlined in
section 2.3
, below.
2.1 Common Misconceptions
Users should be aware of the following characteristics of the PippinHT system. These
are part of normal operation of the system, but may seem counterintuitive at first.
2.1.1 Narrowest is not always the best.
The PippinHT can produce very narrow size distributions from sheared genomic
DNA. However, narrower size distributions will necessarily mean that a smaller
fraction of the input DNA will be recovered. Users should broaden their collection
ranges if the default tight settings do not produce enough DNA for their
application.
Figure 2.1
. A comparison of the relative sample yield between tight and broader range
size selections of DNA.
