RAPID Lite

ELANE

Click on each title for more information

I. DNA amplification

Kit
GE:illustra GenomiPhi V2 DNA Amplification Kit

II. PCR

Primer details

Exon No.	Forward sequence	Reverse sequence	Expected PCR product size (bp)	Genomic regions amplified by ePCR with the primer
1	CGAGCCAATCCAGCGTCTTGTC	TGGCTTCACCGCTCAGAACCTC	406bp	chr19:852151+852556
2	AGGTCTGTCTGTGCCTTGGAG	CTGAGGGCGAAGGTGCTC	542bp	chr19:852634+853175
3	CTCGAGCACCTTCGCCCTCAG	CCCGTTTCACAGAGGTGCAGAC	414bp	chr19:853155+853568
4	GAACCACAGTGGAACCTGAGATG	GTCTAGCCACGGTGCCTGTTG	409bp	chr19:855461+855869
5	CCCTAGGAGGGACTTCCCAACCCTG	CACCACGCCCGACCTACTGACC	442bp	chr19:855869+856310

III. Sequencing reaction

Kit
BigDye® Terminator v3.1 Cycle Sequencing Kit
Primer details

Exon No.	Forward sequence	Reverse sequence	Expected PCR product size (bp)	Genomic regions amplified by ePCR with the primer
1	CGAGCCAATCCAGCGTCTTGTC	TGGCTTCACCGCTCAGAACCTC	406bp	chr19:852151+852556
2	TTCCTGGTGGGGGATCCAGA	CCCACCTCACAGACCGGGA	305bp	chr19:852788+853092
3	CTCGAGCACCTTCGCCCTCAG	CCCGTTTCACAGAGGTGCAGAC	414bp	chr19:853155+853568
4	GAACCACAGTGGAACCTGAGATG	GTCTAGCCACGGTGCCTGTTG	409bp	chr19:855461+855869
5	CCCTAGGAGGGACTTCCCAACCCTG	CACCACGCCCGACCTACTGACC	442bp	chr19:855869+856310

IV. Ethanol precipitation

V. Sequencing analysis

5xPrimeSTAR GXL Buffer	2 µL
2.5mM dNTP	0.8 µL
5pmol/µL Fw primer	1 µL
5pmol/µL Rev primer	1 µL
PrimeSTAR GXL DNA Polymerase	0.2 µL
Genomiphi product	0.5 µL
Sterilized distilled water	4.5 µL