Methodologies for In Vitro Cloning of Small RNAs and Application for Plant Genome(s)

Table 1

Examples of Roche (454) fusion primer sequences and a set of simple bar-coded Roche (454) fusion primer sequences based upon the 𝟑^′ and 𝟓^′ linkers in the IDT miRCat Small RNA Cloning Kit.

miRCat linker-specific PCR primers:
Forward	5^′-TGGAATTCTCGGGCACC-3^′
Reverse	5^′-GATTGATGGTGCCTACAG -3^′
Roche (454) fusion primers:
Forward	𝟓^′-GCCTCCCTCGCGCCATCAGTGGAATTCTCGGGCACC-𝟑^′
adaptor A
Reverse	𝟓^′-GCCTTGCCAGCCCGCTCAGGATTGATGGTGCCTACAG-𝟑^′
adaptor B
Simple fusion primer bar-coding scheme (6 of 256 possible sequences):
5^′-GCCTCCCTCGCGCCATCAGGTACTGGAATTCTCGGGCACC-3^′
5^′-GCCTCCCTCGCGCCATCAGCGATTGGAATTCTCGGGCACC-3^′
5^′-GCCTCCCTCGCGCCATCAGTCGATGGAATTCTCGGGCACC-3^′
5^′-GCCTCCCTCGCGCCATCAGATGCTGGAATTCTCGGGCACC-3^′
5^′-GCCTCCCTCGCGCCATCAGCCTCTGGAATTCTCGGGCACC-3^′
5^′-GCCTCCCTCGCGCCATCAGTGGATGGAATTCTCGGGCACC-3^′