Advances in Hematology

Research Article

Development and Characterization of Anti-Nitr9 Antibodies

Table 1

Oligonucleotide primer sequences.


Purpose	Primer sequence

Reverse transcriptase—PCR: nitr9	GGATTTTTGGACTTTTCTGTC
	TCCACATGCGGTAACTGTAC
Reverse transcriptase—PCR: β-actin	GGTATGGAATCTTGCGGTATCCAC
	ATGGGCCAGACTCATCGTACTCCT
TaqMan Q-PCR: nitr9 (probe = CAAGGTTTGGAAAAGCAC)	GTCAAAGGGACAAGGCTGATAGTT
	GTTCAAAACAGTGCATGTAAGACTCA
TaqMan Q-PCR: β-actin (probe = CCCATGCCATCCTGC)	CCATCTATGAGGGTTACGCTCTTC
	AGGATCTTCATCAGGTAGTCTGTCA
Amplify nitr9 I domain for bacterial expression construct	A TGGAAAAGCACACTGTAGTA^a
	TTATTTAGAGCCATTCCTGTCC^b
Amplify nitr9L for FLAG-tagged expression cassette	CACCCAAATGCACCACCTGTGTTTGTTAAAC^c
	gactgcggccgcTTACTGCTGGTTAGAAAC^d
Amplify nitr9S for FLAG-tagged expression cassette	CACCCAAATGCACCACCTGTG^c
	gactgcggccgcTTACTGCTGGTTAGAAAC^d
Amplify nitr9SS for FLAG-tagged expression cassette	CATGATTTAATTCCATCCCA^c
	gactgcggccgcTTACTGCTGGTTAGAAAC^d
Amplify wild type nitr9L, nitr9S and nitr9SS for expression cassettes	gatcggatccgacATGATCAACTTTTGGATTT^e
	gatcgaattcTTACTGCTGGTTAGAAACCGAG^f

An artificial start codon is underlined.
An artificial stop codon is bold.
These primers are designed for blunt PCR cloning into the EcoRV site of pLF.
Overhang (5′) sequences are in lower case text and include a Not I site for cloning into pLF.
Overhang (5′) sequences are in lower case text and include a BamHI site for cloning into pcDNA3.
Overhang (3′) sequences are in lower case text and include an EcoRI site for cloning into pcDNA3.