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User:DrJones1935/1 August 2012
From 2012.igem.org
I. Order Beta Homolog Primers from Keck
- Primers were ordered with the name/sequence in the following table:
| Name | Tm (Tm overhang/Tm anneal) | Length (bp) | Sequence (overhangANNEAL) |
|---|---|---|---|
| pT5_F1 | 76.37 | 92 | 5'-CTAGTCTAGATTTGCTTTGTGAGCGGATAACAATTATAATAGATTCAATTGTGAGCGGATAACAATTACTAGAGAAAGAGGAGAAAGGTACC-3' |
| pT5_R1 | 76.37 | 92 | 5'-GGTACCTTTCTCCTCTTTCTCTAGTAATTGTTATCCGCTCACAATTGAATCTATTATAATTGTTATCCGCTCACAAAGCAAATCTAGACTAG-3' |
| pUni_F | 66.89 | 32 | 5'-CTAGTCTAGATTTGCTTTGTGAGCGGATAACA-3' |
| pUni_R2 | 64.52 | 32 | 5'-GGTACCTTTCTCCTCTTTCTCTAGTAATTGTT-3' |
| p40_F | 72.13 (64.83/55.53) | 56 (35/21) | 5'-gataacaattactagagaaagaggagaaaggtaccATGGCAACTAAAAAACAAGAG-3' |
| p40_R | 66.39 (25.28/58.42) | 32 (10/22) | 5'-gcttactagtCTATTCATTTGTTTCCCCTCCT-3' |
| p41_F | 73.76 (64.83/58.35) | 54 (35/19) | 5'-gataacaattactagagaaagaggagaaaggtaccATGGCTGAAAATGCTGTCA-3' |
| p41_R* | 68.49 (25.28/59.97) | 25 (10/15) | 5'-gcttactagtTCATGCGTTGGGCCC-3' |
| p42_F | 72.44 (64.83/55.76) | 55 (35/20) | 5'-gataacaattactagagaaagaggagaaaggtaccATGTCAACTAACGACGAATT-3' |
| p42_R | 66.56 (25.28/59.83) | 33 (10/23) | 5'-gcttactagtTTAGATCATTGACCCTTGAACCT-3' |
| p43_F | 72.34 (64.83/56.43) | 57 (35/22) | 5'-gataacaattactagagaaagaggagaaaggtaccATGGCAAATGAATTAGGAATCT-3' |
| p43_R | 66.53 (25.28/58.73) | 30 (10/20) | 5'-gcttactagtTTAGAATCCCTCCAAAGGCT-3' |
| p44_F | 72.07 (64.83/57.21) | 59 (35/24) | 5'-gataacaattactagagaaagaggagaaaggtaccATGGGGAATGAATTAATAGTAAGC-3' |
| p44_R | 67.14 (25.54/58.91) | 28 (10/18) | 5'-gcAtactagtTCAGAAAGGGTAAGGGCC-3' |
| p45_F | 72.09 (64.83/55.29) | 56 (35/21) | 5'-gataacaattactagagaaagaggagaaaggtaccATGGAAAAACCAAAGCTAATC-3' |
| p45_R | 66.46 (25.28/58.33) | 31 (10/21) | 5'-gcttactagtCTAAGAAGCTAAAGGCTGTGT-3' |
| p46_F | 73.48 (64.83/57.67) | 54 (35/19) | 5'-gataacaattactagagaaagaggagaaaggtaccATGAGCACAGCAGTACAAA-3' |
| p46_R* | 66.50 (25.28/61.33) | 40 (10/30) | 5'-gcttactagtTTATGATGCCTTTTTCCTTAAAAAATCAGA-3' |
| p47_F | 72.72 (64.83/55.62) | 54 (35/19) | 5'-gataacaattactagagaaagaggagaaaggtaccATGGCAACACAAAAAGTTG-3' |
| p47_R* | 66.21 (25.28/60.97) | 40 (10/30) | 5'-gcttactagtTTAAGCCTTATCCTGATTAGTTTCTTTTTC-3' |
| p48_F | 70.19 (64.83/55.02) | 63 (35/28) | 5'-gataacaattactagagaaagaggagaaaggtaccATGACTGAAAATAATAAATTACAAACTA-3' |
| p48_R | 66.05 (25.28/58.60) | 31 (10/21) | 5'-gcttactagtTTAAAATGGCTCTTCTTCGCT-3' |
| p49_F | 73.42 (64.83/57.24) | 54 (35/19) | 5'-gataacaattactagagaaagaggagaaaggtaccATGACTAAGCAACCACCAA-3' |
| p49_R* | 66.40 (25.28/61.07) | 39 (10/29) | 5'-gcttactagtTTATTCCTCTGAATTATCGATTACACTGT-3' |
| p50_F | 73.68 (64.83/56.95) | 52 (35/17) | 5'-gataacaattactagagaaagaggagaaaggtaccATGAGTACTGCACTCGC-3' |
| p50_R | 67.27 (25.28/58.80) | 27 (10/17) | 5'-gcttactagtTCATGCTGCCACCTTCT-3' |
| pExo_R | 66.38 (25.28/57.10) | 39 (10/16) | 5'-gcAtactagtTCATCGCCATTGCTCC-3' |
- 1 Ordered separately at 200 nmol scale,
Gel purifiedcassette purified - 2 Ordered separately due to 24 primer limit
- See Project Page for more info
II. Check Sequence Data from 26 Jul 2012
Results look favorable. Files:
