Bio-Rad CHEF Mapper® XA System User Manual

Page 47

Fig. 7.2. Separation and identification of fragments larger than 1 megabase. The samples in
Figure 7.1 were separated in 0.8% agarose for 108 hr in 1x TAE at 14 °C, 2 V/cm. Switch time was 35
min. A. Ethidium bromide stained gel.

S. cerevisiae chromosomes are on the far left, S. pombe on the

right, with sizes indicated in megabases. Lane 1, Nru l; Lane 2, Not 1. B. Autoradiogram after
hybridization. Hybridization was as described in Figure 7.1.

Fig. 7.3. Importance of switch interval for resolving fragments. Human DNA was digested with Mlu
I for 6 hr and separated with different switch times or other pulsed field parameters. After membrane
separation, DNA was transferred to Zeta-Probe membrane as described in Figure 7.1. A. 1% gel elec-
trophoresed for 36 hr at 6 V/cm with a switch ramp from 30 sec to 60 sec. B. 1% gel electrophoresed
36 hr at 6 V/cm with a ramp from 95 sec to 115 sec. C. 0.8% gel electrophoresed 108 hr at 2 V/cm
with a switch time of 35 min. Sizes indicate the position of the yeast markers included in each gel.

1 2

2.2-

1.6-

1.1

-5.7

-4.7

-3.5

6 0 0 -

5 5 0 -

4 4 0 -

2 1 5 -

1 1 2 5 -

9 6 0 -

8 3 6 -

5 5 0 -
4 6 0 -

2 1 5 -

5 7 0 0 -

4 7 0 0 -

3 5 0 0 -

2 2 0 0 -
1 1 2 5 -

2 1 5 -