Materials
Name | Company | Catalog Number | Comments |
Primer F2 | Integrated DNA Technologies | TGACTAACTAGGAGGAATAAATG GCTAAAATGAGAATAT |
|
Primer R2 | ibid | CATTATTCCCTCCAGGTACTAAAA CAATTCATCCAGT |
|
Primer F2p | ibid | GATAAACCCAGCGAACCATTTGA | |
Primer P1 | ibid | GCTTATATACCTTAGCAGGAGACA | |
Primer P2 | ibid | GTATGACATTGCCTTCTGCGTCC | |
Primer 5' end_R1_seq | ibid | GCCATTTATTCCTCCTAGTTAGTCA | |
Primer 5' end_F3_seq | ibid | GTTTTAGTACCTGGAGGGAATAATG | |
Primer 5' end_R1p_seq | ibid | CCTCAAATGGTTCGCTGGGTTTATC | |
CSP | Synprep | Sequence:NH2-DLRGVPNPWGWIFGR-COOH | |
DNA Polymerase | Invitrogen | 11304-102 | Platinum Taq DNA Polymerase High Fidelity |
EcoRI | New England Biolabs Inc | R0101S | Restriction enzyme |
Agar | American Bioanalytical | AB01185 | |
Agarose | Promega | V3125 | |
BHI | BD Biosciences | 237500 | Brain Heart Infusion |
Horse serum | Fisher Scientific | SH3007403 | Horse serum |
Km | Fisher Scientific | BP906-5 | Kanamycin |
TH broth | BD Biosciences | 249240 | Todd Hewitt broth |
PureLink 96 | Invitrogen | K310096 | PureLink 96 PCR purification kit |
QIAquick | Qiagen | 28106 | QIAquick PCR purification kit |
Filter | Corning | 431117 | 0.22 μm polystyrene filter |
Anoxomat System | Mart Microbiology b.v. | Anoxomat Mark II | |
Benchtop centrifuge | Thermo Scientific | 75004377 | Sorvall Legend RT Plus microplate rotor |
Gel Documentation | UVP LLC | BioDoc-It 210 | |
Incubator | Fisher Scientific | 11-690-625D | Isotemp |
Labnet's Gel XL | Labnet | E0160 | Labnet's Gel XL |
Microcentrifuge | Eppendorf | 22621408 | Microcentrifuge 5415 R |
Multichannel pipettes | Thermo Scientfic | 4661040, 4661020 | Finnpipette F1 |
Thermal Cycler | Applied Biosystems Inc. | N805-0200 | GeneAmp PCR system 9700 |
References
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- Pakula, R., Walczak, W. On the nature of competence of transformable streptococci. J. Gen. Microbiol. 31, 125-133 (1963).
- Datsenko, K. A., Wanner, B. L. One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products. Proc. Natl. Acad. Sci. U.S.A. 97, 6640-6645 (2000).
- Xu, P., et al. Genome-wide essential gene identification in Streptococcus sanguinis. Sci. Rep. 1, (2011).