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We performed 5' and 3' RACE experiments using 5' RACE and 3' RACE systems (Invitrogen) according to the manufacturer's protocol.
Subsequently, we performed 3' RACE experiments on P1-rr4B2 total RNA extracted from silk and one primer binding (p2 race 5'-3, see Table 1) to exon 3.
Two cDNA fragments were obtained in initial 5′ RACE experiments.
Rapid amplification of 5′ complementary DNA ends (5′ RACE) experiments employing the test plasmid pEZT7-EGFP revealed that transcripts indeed initiated at pT7.
RACE experiments with two different primers (T10F006553E1 and T10F006553F9) were conducted.
The RACE experiments resulted in two different sxtA - like transcript families.
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This TSS was confirmed by 5'-RACE experiments in 4 out of 6 tissue samples supporting our prediction (primer T10F0065AF50) (for full details on 5'-RACE experiments see Online Supporting Materials for [5]).
We then performed 5'-RACE experiments using total RNA isolated from mutant strains that lack the CRP-cAMP and GalR transcription factors (Fig. 3).
Since 5'-RACE experiments and cDNA library screenings failed to deliver the missing diaphanous sequences, we decided to perform a genome walking approach.
5'-RACE experiments under several conditions were performed, without success.
OKT and KT performed 5'-RACE experiments and analyzed the signaling sequences.
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