Mu-active line of ChinaMu
The original seed stocks 330I (a1-mum2; A2 C1 C2 MuDR R1) containing active Mu were obtained from Maize Genetics Cooperation Stock Center. To identify the linage of 330I, pair-wise distance among all 3241 accessions was calculated based on the p-distance model, and a neighbor-joining (NJ) method (Saitou and Nei, 1987) was used to construct the phylogenetic tree. The result of the phylogenetic tree suggests that the genetic distance between Mu-active line (330I) and W22 is closest (0.24).
PCR confirmation of Mu insertions.
1. Generally we provide 30 F3-generation seeds for each insertion site. To confirm presence of the Mu insertions in a gene of interest, genomic DNA must be extracted from individual plants and analyzed by PCR using gene-specific primers.
2. upstream and one downstream of the insert site), and these will need to be compatible with a Mu TIR specific primer Mu67 (5’-GAAGCCAACGCCAWCGCCTCYATTTCGTCGAAT).
Example:
Figure 1. Primer design for Mu insertion located in the exon region of Zm00001d022184.
‘F’ and ’R’ represent the primers located on both sides of the insertion, and Mu67 represents the Mu-TIR specific primer.
Figure 2. PCR validation for Mu insertions inheritability
‘+/+’, ‘+/-’, ‘-/-’ represent no insertion, heterozygous insertion, and homozygous insertion respectively.
3. We have carried out PCR validation on the inheritability of Mu insertions. This work is still ongoing and will continue to be updated. (These sites are based on the B73 V4 genome)
The attachment is the identified insertion site and related primer information.
If you have any questions, please contact us.
Note: Please be kindly advised that we are NOT responsible for guarantee 100% accuracy of all the insertions in our stocks.
