2X PCR buffer 6/30/2016

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PCR using--- as template, T7-F and T7-R as primers

2X buffer with 1.5 mM Mgcl2  final conc
 Control
 2.5U NEB Taq
1
 0.5U Taq
2
 1U Taq
3
 2.5U Taq
4
 5U Taq
2X buffer with 2.5 mM Mgcl2 final conc
 Control
 2.5U NEB Taq
5
 0.5U Taq
6
 1U Taq
7
 2.5U Taq
8
 5U Taq

20 cycle

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25 cycle

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30 cycle

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35 cycle

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6/30/2016

template 528 pLVX-IRES-EGFP， IRES-Xho1-F +  IRES-Mlu1-R， product 1.3 kb 

ctcgag TACGTAAATTCCGCCCCTCTCCCTAACGTT

ACTGGCCGAA

GCCGCTTGGAATAAGGCCGGTGTGCGTTTGTCTATATGTTATTTTCCACCATATTGCCGTCTTTTGGCAATGTGAGGGCCCGGAAACCTGGCCCTGTCTTC
TTGACGAGCATTCCTAGGGGTCTTTCCCCTCTCGCCAAAGGAATGCAAGGTCTGTTGAATGTCGTGAAGGAAGCAGTTCCTC
TGGAAGCTTCTTGAAGACAAACAACGTCTGTAGCGACCCTTTGCAGGCAGCGGAACCCCCCACCTGGCGACAGGTGCCTCTG
CGGCCAAAAGCCACGTGTATAAGATACACCTGCAAAGGCGGCACAACCCCAGTGCCACGTTGTGAGTTGGATAGTTGTGGAA
AGAGTCAAATGGCTCTCCTCAAGCGTATTCAACAAGGGGCTGAAGGATGCCCAGAA
KpnI    
GGTACC
CCATTGTATGGGATCTGATC
TGGGGCCTCGGTGCACATGCTTTACATGTGTTTAGTCGAGGTTAAAAAAACGTCTAGGCCCCCCGAACCACGGGGACGTGGT
TTTCCTTTGAAAAACACGATGATAATATGGCCACAACC

EGFP
ATGGTGAGCAAGGGCGAGGAGCTGTTCACCGGGGTGGTGCCCAT
CCTGGTCGAGCTGGACGGCGACGTAAACGGCCACAAGTTCAGCGTGTCCGGCGAGGGCGAGGGCGATGCCACCTACGGCAAG
CTGACCCTGAAGTTCATCTGCACCACCGGCAAGCTGCCCGTGCCCTGGCCCACCCTCGTGACCACCCTGACCTACGGCGTGC
AGTGCTTCAGCCGCTACCCCGACCACATGAAGCAGCACGACTTCTTCAAGTCCGCCATGCCCGAAGGCTACGTCCAGGAGCG
CACCATCTTCTTCAAGGACGACGGCAACTACAAGACCCGCGCCGAGGTGAAGTTCGAGGGCGACACCCTGGTGAACCGCATC
GAGCTGAAGGGCATCGACTTCAAGGAGGACGGCAACATCCTGGGGCACAAGCTGGAGTACAACTACAACAGCCACAACGTCT
ATATCATGGCCGACAAGCAGAAGAACGGCATCAAGGTGAACTTCAAGATCCGCCACAACATCGAGGACGGCAGCGTGCAGCT
CGCCGACCACTACCAGCAGAACACCCCCATCGGCGACGGCCCCGTGCTGCTGCCCGACAACCACTACCTGAGCACCCAGTCC
GCCCTGAGCAAAGACCCCAACGAGAAGCGCGATCACATGGTCCTGCTGGAGTTCGTGACCGCCGCCGGGATCACTCTCGGCA
TGGACGAGCTGTACAAGTAA

ACGCGT

PCR primers 

IRES-Xho1-F   2011_12_22    H10  ATCGCACTCGAGTACGTAAATTCCGCCCCTCTC
        Xho1
atcgca ctcgag TACGTAAATTCCGCCCCTCTC


IRES-Mlu1-R 2011_12_22    H11     GATGCGTACGCGTTTACTTGTACAGCTCGTCCATG
(CATGGACGAGCTGTACAAGTAA ACGCGT acgcatc)
gatgcgtACGCGTTTACTTGTACAGCTCGTCCATG
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