Kidd (SLC14A1 rs1058396) Genotyping Performed by PCR-Restriction Fragment Length Polymorphism Using a General-Purpose Agarose Gel and Lithium Borate Buffer / 대한수혈학회지
Korean Journal of Blood Transfusion
; : 154-160, 2022.
Article
in English
| WPRIM (Western Pacific)
| ID: wpr-967997
Responsible library:
WPRO
ABSTRACT
Background@#Kidd (SLC14A1 rs1058396) genotyping is an important test to prevent delayed hemolytic transfusion reactions and hemolytic disease of the fetus or newborn. The PCR-restriction fragment length polymorphism (RFLP) technique using the MnlI restriction enzyme is not used widely because of the need for a polyacrylamide gel.PCR-RFLP was performed by electrophoresis with general-purpose agarose gel, and lithium borate buffer (LBB) was developed as a replacement for polyacrylamide gel. @*Methods@#Seventy-two venous blood samples were collected randomly and used in this study. A 3% agarose gel containing 1 µg/mL of ethidium bromide and 1 mM LBB, and a high-voltage (300 V) were used to separate the short-length restriction fragments (72 bp, 51 bp, 36 bp, and 21 bp). The PCR-RFLP results were confirmed by PCR-direct sequencing. @*Results@#Target restriction fragments could be easily discriminated. The results obtained with the PCR-RFLP were completely in agreement with the results of PCR-direct sequencing. @*Conclusion@#The PCR-RFLP using a general-purpose agarose gel and LBB is an accurate and reliable assay for Kidd genotyping.
Full text:
Available
Database:
WPRIM (Western Pacific)
Language:
English
Journal:
Korean Journal of Blood Transfusion
Year:
2022
Document type:
Article