Page 266 - Plant Canada 2024 Proceeding

PLANT CANADA 2024

[P126] DISCOVERY OF YIELD QTL IN CANADIAN SPRING WHEAT. Santosh Kumar , Jasdeep Kaur ,
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Clare Workman , and Kirby Nilsen . Brandon Research and Development Centre, Agriculture and Agri-
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Food Canada, 2701 Grand Valley Road, Brandon, MB, Canada, R7A 5Y3
Correspondence to: santosh.kumar@agr.gc.ca

The northern Prairie region of Canada has a shorter growing season, which impedes the yield potential
due to insufficient growing degree days. Yield is the most important factor in the farm scale adoption and
profitability of a new variety. Among yield contributing factors, days to maturity and yield are negatively
corelated. To understand the yield contributing factors, a doubled haploid population was developed from
a cross between AAC Wheatland (mid-season) and breeding line PT485 (early maturing). Both AAC
Wheatland and PT485 are semi-dwarf premium quality wheat varieties but differ in AAC Wheatland being
Orange Blossom Wheat Midge tolerant, which provides yield advantage. The DH population was grown
as yield plots at Brandon, MB and Saskatoon, SK. The data on yield components (height, maturity, yield,
test weight and thousand kernel weight) were recorded. Genotyping was performed on the mapping
population using Affymetrix 25K SNP Chip through SGS North America. The linkage map was
constructed using MSTmap resulting in 36 linkage groups using 4552 polymorphic loci. The QTL analysis
was performed on Qgene software. A yield QTL was detected in both environments on chromosome 7.
The closest SNPs AX-158544327 and TGWA25K-TG0227 are at 7.1 cM and 1 cM from the QTL peak
respectively suggesting close linkage. The poster will describe the potential QTL for yield and the
underlying genomic loci with genes of interest.

[P127] MAGNETIC BEAD BASED PLASMID ISOLATION PROTOCOL FOR HIGH-YIELD
SEQUENCING GRADE PLASMID DNA. Ankita Talla, Sneha Thakur, Lalitha S., Vishal Mane, Radha
Hariharan, Sujata Hajra, Kavita Khadke, and Rajas Warke. HiMedia Laboratories Pvt. Ltd.
Correspondence to: kavita.khadke@himedialabs.com

The recombinant DNA technology has demonstrated unique impacts in bringing advancement in human
life. This technology has various applications and potential to deal with important aspects of life, for
instance, improving health, enhancing food resources, and resistance to various environmental effects.
Particularly in agriculture, the genetically modified plants have increased resistance to harmful agents,
enhanced product yield, and shown increased adaptability for better survival.

The basic steps of molecular cloning includes vector digestion, insert preparation, ligation, transformation
and Clone selection. Hence, Plasmid vectors and clones with high yield and good purity are always in
high demand. In HiMedia, we have indigenously developed one automated magnetic bead-based system
which consistently isolates recombinant plasmids having high yield and purity.

This study signifies using Insta NX Automated extraction platform for plasmid DNA extraction from small
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volume of cultures. HiPurA Pre- filled Cartridges for Plasmid DNA Extraction MB508PC16 along with
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Insta NX Mag16 Plus provides the fastest and easiest way to purify plasmids of varying size under
affordable cost and less time. This extraction method is found to be most efficient and suitable for
applications in PCR, library screening, sequencing, etc. The DNA purification procedure comprises of
three steps viz. adsorption of DNA to the magnetic beads, removal of residual contaminants and elution
of pure plasmid DNA. HiMedia’s Insta NX Mag16 Plus can be used for processing small amount of
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samples volumes (from 500µl to 5ml) for plasmid DNA extraction. An average yield upto 50ng/µl and
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purity (A260/ A280) in range of 1.7-1.9 is achieved by MB508PC16 - HiPurA Pre- filled Cartridges for
Plasmid DNA Extraction kits.

The number of steps involved in manual plasmid DNA extraction are minimized with the help HiMedia’s
Magnetic Automated extractor and HiPurA Pre-filled kits. The HiPurA Pre- filled Cartridges for Plasmid
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DNA Extraction kit provide a high quality and quantity of plasmid DNA required for Recombinant Protein
expression, PCR analysis, Library preparation followed by Sequencing and other downstream molecular
applications.

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