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Pluripotency Check PCR Kit

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Clontech
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Pluripotency Check PCR Kit
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Pluripotency Check PCR Primer Set
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16 x 25 rxns
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Validate the pluripotent status of your stem cell or iPS cell culture using a simple PCR kit
  • 16 prevalidated PCR primer mixes
  • Complete kit with RNA extraction, reverse transcription, and PCR reagents
  • Confirm your iPS cells by differentiating between endogenous and exogenous gene expression

Confirming that your mouse stem cells or iPS cells remain pluripotent is a critical step in stem cell research. Now, you can quickly validate the pluripotent status of your mES cells via PCR, using a simple, prevalidated primer set (1) or a start-to-finish kit.



Figure 1. Confirming the pluripotent status of an mES culture. Total RNA was extracted from ES-E14TG2a mouse embryonic stem cells (mES), followed by cDNA synthesis and amplification according to the protocol outlined in PT5086-2. M: 100 bp DNA Marker. Lane1, Total Oct3/4. Lane 2: Endogenous Oct3/4. Lane 3: Total Nanog. Lane 4: Endogenous Nanog. Lane 5: Total c-Myc. Lane 6: Endogenous c-Myc. Lane 7: Total Sox2. Lane 8: Endogenous Sox2. Lane 9: Total Klf4. Lane 10: Ecat1. Lane 11: ERas. Lane 12: Esg1. Lane 13: Rex1. Lane 14: GAPDH (control). Lane 15: ¥â-Actin (control). Endogenous Klf4, data not shown.

The Pluripotency Check PCR Kit contains all the necessary reagents to confirm that your mouse stem or iPS cells are indeed pluripotent. It includes 16 prevalidated forward and reverse primer mixes, based on nine genes expressed in pluripotent stem cells and two control genes (1; Table I). The kit includes everything for start-to-finish confirmation of your cells¡¯ pluripotency status, using any combination of primer sets, or the entire panel.

The Pluripotency Check PCR Primer Set consists of 16 prevalidated primer mixes based on nine genes expressed in pluripotent stem cells, plus two control genes. The different amplicon sizes allow you to distinguish between engogenous and exogenous expression.

Table I. Pluripotency Check PCR Primers
Primer Set Amplicon Size Primer Set Amplicon Size
Oct3/4 (Total)1 485 bp Klf4(Total)1 739 bp
Oct3/4 (Endogenous)1 224 bp Klf4 (Endogenous) 206 bp
Nanog (Total)1 364 bp Ecat11 164 bp
Nanog (Endogenous)1 223 bp ERas1 210 bp
Sox2 (Total)1 193 bp Esg11 376 bp
Sox2 (Endogenous)1 297 bp Rex11 287 bp
c-Myc (Total)1 228 bp ©¬-Actin (Control) 603 bp
c-Myc (Endogenous)1 170 bp GAPDH (Control) 568 bp
References
1. Takahashi, K., and Yamanaka, S. (2006) Cell 126(4):663-676.
Components
- Pluripotency Check PCR Primer Set (16 x 25 rxns)
- NucleoSpin RNA II (10 preps)
- SMART MMLV Reverse Transcriptase (8,000 units)
- TITANIUM Taq PCR Kit (30 rxns)
The Pluripotency Check PCR Primer Set is also sold separately.
Storage Conditions
Store the NucleoSpin RNA II Kit at room temperature.
Store all other components at -20¡ÆC.

Keyword :

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