Table 1 Table of primers: primers used in the pre-amplification, LAMP, RPA, qPCR, and HRM reactions. Primer’s sequences for male detection. The following primers are from 9 and 15: MADC2 Fwd and MADC2 Rev. The other primers listed are novel designs that target the same amplicon region or a nearby loci to gain the same discrimination between males and non-males. The first column designates the type of end-point assay to be used that include loop-mediated isothermal amplification (LAMP), recombinase polymerase amplification (RPA), quantitative polymerase chain reaction (qPCR), and high-resolution melting (HRM). All sequences are provided in a 5′ to 3′ orientation
From: High-throughput methods to identify male Cannabis sativa using various genotyping methods
Assay | Primer name | Primer sequence (5′ −> 3′) |
|---|---|---|
LAMP | F3 | AAATGCCAGGTTCGATCA |
B3 | TCTTAGGTGAATTACCTGTACG | |
FIP | TGCATTAGCTCAAAACTAATTGGGT-GAACTGCATATCCATATATTCTCC | |
BIP | GCTATTCCTGAAACTTTTGAAACGT-CAAGTCTTGCCTTGAAGGA | |
LF | TAGATCTTGCGAATTGATAGGGGT | |
LB | GCAAAGAAAGAACGTTTGGTCATCT | |
RPA | TwstoMndo3 Fwd | CTACTATGGAGTGCTAGGGGCAGTTAATTGAG |
TwstoMndo5 Rev | TCCACTGAAAAATATCGTCTTAGGTGAATTACCTG | |
qPCR | TC Universal CS Fwd | TGTCCTACATATCTCAAGTCCCATTT |
TC Universal CS Rev | AAGGGTAGCTCCGGCTTCAA | |
TC Universal CS TaqMan probe | /5HEX/TA+GTAGA+CTTGA+GAAA+CATGC/3BHQ_1/ | |
MADC2 Fwd | GTGACGTAGGTAGAGTTG | |
MADC2 Rev | GTGACGTAGGCTATGAGAG | |
MADC2 TaqMan probe | /56-FAM/CT+AAT+GCA+GG+CTATT+CC/3BHQ_1/ | |
HRM | Choco Mando Fwd | CTGCATATCCATATATTCTC |
Choco Mando Rev | GTGACGTAGGCTATGAGAG | |
Break 1 Fwd | TGGTCCGCAAAGAAAGAACGTTTG | |
Plus 9 Rev | GATCAAACCTATTTATGGCATTTAC | |
MADC2 Fwd | GTGACGTAGGTAGAGTTG | |
MADC2 Rev | GTGACGTAGGCTATGAGAG | |
−403 Fwd | GAGAAATTTCTTTGATTAGCAAAG | |
−237 Rev | GGACTATTTATAGTCGCAAGGATG |