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)


Vu et al. BMC Genomics 2010 11:301   doi:10.1186/1471-2164-11-301

Open Data