. Author manuscript; available in PMC: 2013 Jul 26.

Published in final edited form as: Nat Protoc. 2012 Feb 2;7(2):374–393. doi: 10.1038/nprot.2011.446

TABLE 1.

Primers.

Primer	Use	Working concentration	Sequence
5′miR30-XhoI	PCR cloning	10 µM	5′-TACAATACTCGAGAAGGTATATTGCTGTTGACAGTGAGCG-3′
3′miR30-EcoRI	PCR cloning	10 µM	5′-ACTTAGAAGAATTCCGAGGCAGTAGGCA-3′
97-mer template	PCR cloning	0.02 ng µl⁻¹	5′-TGCTGTTGACAGTGAGCG-22merSense-TAGTGAAGCCACAGATGT A-22merGuide-TGCCTACTGCCTCGGA-3′
Linker A (5′-phosphorylated), 110 bp	Linker cloning	1 µg µl⁻¹	5′phos-TCGAGAAGGTATATTGCTGTTGACAGTGAGCG-22merSense- TAGTGAAGCCACAGATGTA-22merGuide-TGCCTACTGCCTCGG-3′
Linker B (5′-phosphorylated)	Linker cloning	1 µg µl⁻¹	5′phos-AATTCCGAGGCAGTAGGCA-RC22merGuide-TACATCTGTGG CTTCACTA-RC22merSense-CGCTCACTGTCAACAGCAATATACCTTC-3′
miR30seq	Sequencing	0.5 µM	5′-TGTTTGAATGAGGCTTCAGTAC-3′
shRNA-specific forward	Genotyping	10 µM	See Step 57
RBG-R1	Genotyping	10 µM	5′-GAAAGAACAATCAAGGGTCC-3′
RBG-R2	Genotyping	10 µM	5′-CACCCTGAAAACTTTGCCCC-3′
ColA1 forward	Genotyping	10 µM	5′-AATCATCCCAGGTGCACAGCATTGCGG-3′
ColA1 reverse	Genotyping	10 µM	5′-CTTTGAGGGCTCATGAACCTCCCAGG-3′
SAdpA reverse	Genotyping	10 µM	5′-ATCAAGGAAACCCTGGACTACTGCG-3′

Abbreviations: 5′Phos, the oligonucleotide should be phosphorylated at the 5′ end; 22merSense, unique 22mer sense strand (see Figure 2); 22merGuide, unique 22mer guide strand (see Figure 2); RC22merSense, reverse complement of ‘22merSense’; RC22merGuide, reverse complement of ‘22merGuide’.