MS-PCR primer sequences and reaction conditions
Sequences | 1× Buffer (and additives) | Annealing temperature (°C) | |
---|---|---|---|
M3 | S 5′-GGTTAGTAGTTCGGGTAGGGTTTATC-3′, AS 5′-CCGAATCCTACCCCGACG-3′ | 16.6 mmol/L (NH4)2SO4 | 57 |
67 mmol/L Tris (pH 9.1) | |||
3.0 mmol/L MgCl2 | |||
U3 | S 5′-TTAGTAGTTTGGGTAGGGTTTATT-3′, AS 5′-CCAAATCCTACCCCAACA-3′ | 15 mmol/L (NH4)2SO4 | 57 |
60 mmol/L Tris (pH 8.5) | |||
4.5 mmol/L MgCl2 | |||
M4 | S 5′-GTCGAGAACGCGAGCGATTC-3′, AS 5′-CGACCAATCCAACCGAAACG-3′ | 15 mmol/L (NH4)2SO4 | 55 |
60 mmol/L Tris (pH 9.0) | |||
3.5 mmol/L MgCl2 | |||
150 mmol/L 2-pyrrolidinone | |||
U4 | S 5′-GATGTTGAGAATGTGAGTGATTT-3′, AS 5′-AACCAATCCAACCAAAACA-3′ | 15 mmol/L (NH4)2SO4 | 57 |
60 mmol/L Tris (pH 8.5) | |||
4.5 mmol/L MgCl2 |
Abbreviations: S, sense; AS, antisense.