Difference between revisions of "PCR Primers for Transgenic Mice"

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Line 18: Line 18:
  
 
*Transgene: tetO-G-alpha 12
 
*Transgene: tetO-G-alpha 12
*5´ primer: CGA GAG CGG CAA GTC CAC
+
  *5´ primer: CGA GAG CGG CAA GTC CAC
*3´ primer: GCA GGC TCA GTC GGC ATG TAC
+
  *3´ primer: GCA GGC TCA GTC GGC ATG TAC
*PCR Fragment Size: 245 bp
+
  *PCR Fragment Size: 245 bp
  
Transgene: tetO-G-alpha i2
+
*Transgene: tetO-G-alpha i2
5´ primer: GCA GAT CGA CTT TGC TGA CC
+
  *5´ primer: GCA GAT CGA CTT TGC TGA CC
 +
  *3´ primer: TCT GTC GGC ATG TAC TCC TC
 +
  *PCR Fragment Size: 240 bp
  
3´ primer: TCT GTC GGC ATG TAC TCC TC
+
*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
  
PCR Fragment Size: 240 bp
 
  
Transgene: tetO-G-alpha q
+
*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
  
5´ primer: TTG ACG TCC CCG ACT ACG CG
+
Transactivator Lines
3´ primer: CTT TGG CCC CCT ACA TCG AC
 
  
PCR Fragment Size: 258 bp
+
*Transgene: tTA
 
+
  *5´ primer: GCT GCT TAA TGA GGT CGG
 
+
  *3´ primer: CTC TGC ACC TTG GTG ATC
Transgene: tetO-Ro1/Ro2
+
  *PCR Fragment Size: 507 bp
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
+
*Transgene: MH-tTA
5´ primer: TCA GAC CGA GAT TTC TTC ATC CC
+
  *5´ primer: TCA GAC CGA GAT TTC TTC ATC CC
3´ primer: CTC TGC ACC TTG GTG ATC
+
  *3´ primer: CTC TGC ACC TTG GTG ATC
PCR Fragment Size: ~1,000 bp
+
  *PCR Fragment Size: ~1,000 bp
  
Transgene: MMTV-tTA
+
*Transgene: MMTV-tTA
5´ primer: CTG ATC TGA GCT CTG AGT G
+
  *5´ primer: CTG ATC TGA GCT CTG AGT G
3´ primer: CTC TGC ACC TTG GTG ATC
+
  *3´ primer: CTC TGC ACC TTG GTG ATC
PCR Fragment Size: 595 bp  
+
  *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:'''

Latest revision as of 14:13, 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]