PCR Primers for Transgenic Mice: Difference between revisions
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'''All oligonucleotide sequences are shown as they should be ordered.''' | '''All oligonucleotide sequences are shown as they should be ordered.''' | ||
Tet-Responsive Lines (tetO) | |||
Transgene: tetO-Lac Z | |||
5´ primer: GGC GTG TAC GGT GGG AGG | |||
3´ primer: GTT GGG AAG GGC GAT CGG | |||
PCR Fragment Size: 421 bp | |||
Transgene: tetO-G-alpha 12 | |||
5´ primer: CGA GAG CGG CAA GTC CAC | |||
3´ primer: GCA GGC TCA GTC GGC ATG TAC | |||
PCR Fragment Size: 245 bp | |||
Transgene: tetO-G-alpha i2 | |||
5´ primer: GCA GAT CGA CTT TGC TGA CC | |||
3´ primer: TCT GTC GGC ATG TAC TCC TC | |||
PCR Fragment Size: 240 bp | |||
Transgene: tetO-G-alpha q | |||
5´ primer: TTG ACG TCC CCG ACT ACG CG | |||
3´ primer: CTT TGG CCC CCT ACA TCG AC | |||
PCR Fragment Size: 258 bp | |||
Transgene: tetO-Ro1/Ro2 | |||
5´ primer (Go148): GTT GGG AAG GGC GAT CGG | |||
3´ primer (Go147): GGC GTC TAC GGT GGG AGG | |||
PCR Fragment Size: 178 bp | |||
Transactivator Lines | |||
Transgene: tTA | |||
5´ primer: GCT GCT TAA TGA GGT CGG | |||
3´ primer: CTC TGC ACC TTG GTG ATC | |||
PCR Fragment Size: 507 bp | |||
Transgene: MH-tTA | |||
5´ primer: TCA GAC CGA GAT TTC TTC ATC CC | |||
3´ primer: CTC TGC ACC TTG GTG ATC | |||
PCR Fragment Size: ~1,000 bp | |||
Transgene: MMTV-tTA | |||
5´ primer: CTG ATC TGA GCT CTG AGT G | |||
3´ primer: CTC TGC ACC TTG GTG ATC | |||
PCR Fragment Size: 595 bp | |||
'''Protocols for genotyping the following mice can be found on the Protocol's Page:''' | '''Protocols for genotyping the following mice can be found on the Protocol's Page:''' |
Revision as of 14:08, 29 June 2009
Conklin Lab Transgenic Mice
The Conklin Lab maintains several transgenic lines that utilize the tetracyclinetransactivator system. These lines will be made available on a collaborative basis and as our limited resources allow. (Jackson Lab should have manyof these lines soon).
Genotyping mice
Mice can be genotyped by PCR. After digesting a small piece of mousetail with proteinase K, a small alliquot of sample which contains genomic DNA is analyzed by PCR using transgene specific primers. Full protocol available.
All oligonucleotide sequences are shown as they should be ordered.
Tet-Responsive Lines (tetO)
Transgene: tetO-Lac Z 5´ primer: GGC GTG TAC GGT GGG AGG 3´ primer: GTT GGG AAG GGC GAT CGG PCR Fragment Size: 421 bp
Transgene: tetO-G-alpha 12 5´ primer: CGA GAG CGG CAA GTC CAC 3´ primer: GCA GGC TCA GTC GGC ATG TAC PCR Fragment Size: 245 bp
Transgene: tetO-G-alpha i2 5´ primer: GCA GAT CGA CTT TGC TGA CC
3´ primer: TCT GTC GGC ATG TAC TCC TC
PCR Fragment Size: 240 bp
Transgene: tetO-G-alpha q
5´ primer: TTG ACG TCC CCG ACT ACG CG 3´ primer: CTT TGG CCC CCT ACA TCG AC
PCR Fragment Size: 258 bp
Transgene: tetO-Ro1/Ro2
5´ primer (Go148): GTT GGG AAG GGC GAT CGG
3´ primer (Go147): GGC GTC TAC GGT GGG AGG
PCR Fragment Size: 178 bp
Transactivator Lines Transgene: tTA 5´ primer: GCT GCT TAA TGA GGT CGG 3´ primer: CTC TGC ACC TTG GTG ATC PCR Fragment Size: 507 bp
Transgene: MH-tTA 5´ primer: TCA GAC CGA GAT TTC TTC ATC CC 3´ primer: CTC TGC ACC TTG GTG ATC PCR Fragment Size: ~1,000 bp
Transgene: MMTV-tTA 5´ primer: CTG ATC TGA GCT CTG AGT G 3´ primer: CTC TGC ACC TTG GTG ATC PCR Fragment Size: 595 bp
Protocols for genotyping the following mice can be found on the Protocol's Page:
TetO-Rs1 mice: Protocol to Genotype Rs1 Mice [1]
ColI(2.3)-tTA mice Protocol to Genotype ColI [2] (2.3)-tTA Mice: R. 20080528
NKX2-5-EmGFP (RP11-88L12 NKX2-5 EmGFP reporter mice) Protocol to Genotype NKX2-5 BAC Mice [3]