Table 1.

Comparison of screening BAC library by conventional PCR and by MT-PCR-HRM.

Compared features	Screening by conventional PCR using 3D pooling strategy	Screening by MT-PCR-HRM using 2D pooling strategy
Maximum number of 384-well plates to be pooled in one super pool	10 384-well plates	25 384-well plates
Number of super pools to be tested to identify a positive superpool (N: The total plate number of the BAC library)	N/10	N/25
PCR reactions (PCR/PCR-HRM) needed to identify a positive super pool	N/10	N/25
Reactions needed to identify the plate ID for one positive super pool	10	10
Reactions needed to identify the clone ID from one positive plate	40	18
Total number of reactions needed to get positive BAC clone ID from whole library	N/10 + 50	N/25 + 28
Multiplexing possibility	No	Yes
Checking on agarose gel	Needed	NO
Cost	0.16 £/1 PCR reaction + cost for agarose gel electrophoresis	0.17 £/1 PCR-HRM reaction
Procedure duration to anchor 1 marker	12 h	3 h
Scoring data	Manually from gel photos	Semi-Automatic (figures and summary tables can be exported from HRM rotor system)