A Novel -72 (T→A) β-Promoter Mutation Causing Slightly Elevated HbA2 in a Vietnamese Heterozygote
Table 2
List of primers used to carry out PCR, cloning, site-direct mutagenesis, and sequencing reactions. Sequencing reactions were performed by means of the same primers used for PCR and cloning.
(a)
Primer pairs used for standard PCR and sequencing
Primer code
Sequence (5′ to 3′)
Nucleotide position (#U01317)
Product size
β1(F)
GCCAAGGACAGGTACGGCTGTCATC
61997–62021
706 bp
β2(R)
CCCTTCCTATGACATGAACTTAACCAT
62676–62702
β7(F)
TCCTGATGCTGTTATGGGCAA
62469–62489
923 bp
β8(R)
AAAAGCAGAATGGTAGCTGGA
63371–63391
β9(F)
AAAAACTTTACACAGTCTGCC
62935–62955
966 bp
β10(R)
ATTAGCTGTTTGCAGCCTCA
63881–63900
(b)
Primer pairs used for cloning and sequencing
Primer code
Sequence (5′ to 3′)
Nucleotide position (#U01317)
Product size
β_KpnI(F)
ggtaccATCCAGTTTCTTTTGGTTAACCT
60678–60700
1505 bp
β_XhoI(R)
ctcgagTCTGTTTGAGGTTGCTAGTGAACAC
62158–62182
HS2_BamHI(F)
ggatccTAAGCTTCAGTTTTTCCTTAGT
8485–8506
740 bp
HS2_SalI(R)
gtcgacTAGATCTGACCCCGTATGTGAGCAT
9200–9224
(c)
Primer pairs used for site-direct mutagenesis
Primer code
°Sequence (5′ to 3′)
-87G(F)
CTCACCCTGTGGAGCCACACGCTAGGGTTGGCCAATCTAC
-87G(R)
GTAGATTGGCCAACCCTAGCGTGTGGCTCCACAGGGTGAG
-71T(F)
CTAGGGTTGGCCAATTTACTCCCAGGAGCAGG
-71T(R)
CCTGCTCCTGGGAGTAAATTGGCCAACCCTAG
# = Genbank accession number. Restriction sites are in lowercase. Mutated nucleotides are in boldface and italic. The F and R primer sequences are complementary to each other.
