RNA Detection

9. Because the efficiency of the lysis in this condition is signifi-
   cantly lower than in the condition using detergents and cyclo-
   heximide, a larger amount of starting materials is needed.


10. Harringtonine and 4E1RCat are added to inhibit the transla-
    tion initiation [12].

Acknowledgments

This work was supported by Samsung Science and Technology

Foundation under Project Number SSTF-BA1602-13 (to H.J.)

and by Wellcome Trust Programme Grant (085314/Z/08/Z)

(to C.E.H.).

Eye SC

cre +- +-

INPUT

(1/50 TRAP)

TRAP

0.25

(kb)

0.5

1.0

2.0

5.0

0.25

(kb)

0.5

1.0

2.0

5.0

Fig. 3Expected results. Double strand cDNAs made from TRAPed RNAs from

neonatal mice (postnatal day 0.5). Three eyes or three pairs of superior colliculi

were lysed in ~500μL CHX-lysis buffer. Total 10μL of this lysate was used as

input (upper), and 500μL lysate was TRAPed (lower). Polyadenylated RNA was

made into double strand DNA using the method described by Tang and

colleagues [11]

Isolation of In Vivo Axonal Translatome 93