Primers with degenerate codon and TRIM used for construction of 10 nanobody sub-libraries
| No. | Name | Sequence |
|---|---|---|
| 1 | VHH M13R For | AATTTCACACAGGAAACAGCTATGACCATGATT |
| 2 | FR1-b | CGAGGCGGCACAAGACAA |
| 3 | H1-f | GCCGCCTCGGGGWMTATTTYT####ATGTCGTGGGTACGCCAGGCA |
| 4 | H1-b | TGACACCCACTCCAAGCC |
| 5 | H2-f | GGCTTGGAGTGGGTGTCARSTATTRVT#GGTRSTANTACCWATTATGCCGATTCGGTCAAA |
| 6 | H2-b | ARCCGCGCAATAATAGACAGCGGTATCTTCAG |
| 7-1 (12) | H3-f-1 | GCTGTCTATTATTGCGCGGYT[#=07]YWT#TACTGGGGGCAGGGG |
| 7-2 (14) | H3-f-2 | GCTGTCTATTATTGCGCGGYT[#=09]YWT#TACTGGGGGCAGGGG |
| 7-3 (15) | H3-f-3 | GCTGTCTATTATTGCGCGGYT[#=10]YWT#TACTGGGGGCAGGGG |
| 7-4 (16) | H3-f-4 | GCTGTCTATTATTGCGCGGYT[#=11]YWT#TACTGGGGGCAGGGG |
| 7-5 (17) | H3-f-5 | GCTGTCTATTATTGCGCGGYT[#=12]YWT#TACTGGGGGCAGGGG |
| 7-6 (18) | H3-f-6 | GCTGTCTATTATTGCGCGGYT[#=13]YWT#TACTGGGGGCAGGGG |
| 7-7 (19) | H3-f-7 | GCTGTCTATTATTGCGCGGYT[#=14]YWT#TACTGGGGGCAGGGG |
| 7-8 (20) | H3-f-8 | GCTGTCTATTATTGCGCGGYT[#=15]YWT#TACTGGGGGCAGGGG |
| 7-9 (21) | H3-f-9 | GCTGTCTATTATTGCGCGGYT[#=16]YWT#TACTGGGGGCAGGGG |
| 7-10 (22) | H3-f-10 | GCTGTCTATTATTGCGCGGYT[#=17]YWT#TACTGGGGGCAGGGG |
| 8 | VHH G3P Rev | CTGTATGAGGTTTTGCTAAACAACTTTCAACAGTTT |
“#” denotes random trinucleotides of TRIM. TRIM, trinucleotide-directed mutagenesis.