Effect of DTT and organic reagents on DNA quality and quantity of protein-rich seeds of chickpea (Cicer arietinum L.)

Chickpea is a legume with richness in protein. Therefore, DNA isolation of seeds with proper purity is a great challenge for any molecular studies of this crop due to its abundance in protein and carbohydrates. In this research, a chickpea seeds collection from various seed classes of candidate varieties was tested for genetic purity. Three experiments were planned to find an appropriate protocol for DNA extraction of chickpea seed. SDS based seed extraction buffer used along with, two levels of DTT (Dithiothreitol) and PVP (Polyvinylpyrrolidone) including, ۲% PVP with ۰.۴%, ۰.۸% DTT and applying organic reagents including phenol: chloroform (۲۵:۲۴, v/v) and chloroform: isoamyl alcohol (۲۴:۱, v/v). The data pointed out that applying DTT increase DNA yield that the highest DNA concentration achieved in ۰.۸% DTT ۰, while increasing DTT to ۰.۸% had an adverse effect on DNA quality. Organic reagents along with ۰.۴% DTT presented the best result in terms of DNA quality. Both ۲۶۰/۲۸۰ and ۲۶۰/۲۳۰ ratios were above ۲ and the DNA yield was acceptable for molecular studies. As a result, using DTT can increase DNA yield in chickpea seeds but not great impact on decreasing protein residuals. Exploiting organic reagents with ۰.۴% DTT was efficient to remove protein residual from chickpea seeds.

کلیدواژه ها:

Chickpea ، protein ، DNA extraction ، DTT ، Organic reagent.