microRNA Booster Kit
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The Stemgent microRNA Booster Kit enhances, and simplifies mRNA reprogramming for fast, efficient iPS cell line generation from both normal and patient specific cell lines.The microRNA Booster Kit includes the microRNA Reprogramming Cocktail and B18R Recombinant Protein.
The microRNA-enhanced mRNA Reprogramming method improves upon the first generation mRNA Reprogramming Kit, delivering faster reprogramming kinetics, a streamlined protocol and improved efficacy on hard-to-reprogram or refractory cell types. The microRNA-enhanced method combines Stemgent’s mRNA Reprogramming Kit with the microRNA Booster Kit and the Stemfect™ RNA Transfection Kit to generate expandable primary iPS cell colonies in as little as 8 days. This new method improves kinetics and reduces mRNA dose per transfection resulting in a per well cost reduction of approximately 35% to 45% versus feeder-based mRNA Reprogramming.
mRNA Reprogramming Method
mRNA Reprogramming Method
|Fast Reprogramming Kinetics||Yes||Yes|
|Somatic Cell → iPS Cell||3 weeks||2 weeks|
|Risk of insertional mutagenesis||None||None|
|Screening for Viral Clearance||None||None|
|Screening for DNA vector retention and/or integration||None||None|
|Bio-containment issues associated with viral applications||None||None|
# of wells per kit
(6 well plate format)
(35%-45% per well cost reduction)
|Reprogramming Substrate||Fibroblast feeder layer||Extracellular Matrix|
|Captures hard-to reprogram and refractory cell types||—||Yes|
|Simple, user friendly protocol (overnight transfections)||—||Yes|
- 2 vials - microRNA Reprogramming Cocktail (20 µM) (Cat. No. 05-0036), 35 µL
- B18R Recombinant Protein, Carrier-Free (Cat. No. 03-0017), 50 µg
Storage and Stability
- Store both the microRNA Reprogramming Cocktail and the B18R protein at or below -70 °C.
- Kit components are stable for a minimum of 3 months from date of receipt when stored as directed.
The Stemgent microRNA Booster Kit is functionally validated for successful RNA-based reprogramming of human newborn foreskin fibroblasts. All components of the kit are sterile and have tested negative for mycoplasma.
- Refer to the microRNA-Enhanced mRNA Reprogramming Protocol for directions on use of the Stemgent microRNA Booster Kit.
- Refer to the protocol Generating NuFF-Conditioned Pluriton Medium for instructions on production of the required reprogramming medium.
- Refer to the User Manual: Stemgent mRNA Reprogramming System for additional details on materials and handling.
- RNA Transfection Kit (00-0069)
- mRNA Reprogramming Kit (00-0071)
- NuFF-RQ™ IRR (GSC3002G)
- hPSC XF Medium for Human iPS and ES Cells (01-0005)
- FGF-basic, Human Recombinant (03-0002)
- Stemgent® StainAlive™ TRA-1-60 Antibody (DyLight™ 488), Mouse anti-Human (09-0068)
- Stemgent® StainAlive™ TRA-1-81 Antibody (DyLight™ 488), Mouse anti-Human (09-0069)
- Stemgent® Nanog Antibody (Affinity Purified), Rabbit anti-Mouse/Human (09-0020)
- Stemgent® Oct4 Antibody (Affinity Purified), Rabbit anti-Mouse/Human (09-0023)
- Stemgent® SSEA-4 Antibody (Purified), Mouse anti-Human (09-0006)
- Stemgent® TRA-1-60 Antibody (Purified), Mouse anti-Human (09-0010)
- Stemgent® TRA-1-81 Antibody (Purified), Mouse anti-Human (09-0011)
- mRNA Reprogramming Factors Set: hOKSML (00-0067)
- Burnett LC; LeDuc CA; Sulsona CR; Paull D; Eddira S; Levy B; Salles JP; Tauber M; Driscoll DJ; Egli D; Leibel RL. "Induced pluripotent cells (iPSC) created from skin fibroblasta of patients with Prader-Willi syndrome (PWS) retain the molecular signature of PWS." Stem Cell Res (2016), doi: 10.1016/j.scr.2016.08.008
- Ohlemacher SK; Sridhar A; Xiao Y; Hochstetler AE; Sarfarazi M; Cummins TR; Meyer JS. "Stepwise differentiation of retinal ganglion cells from human pluripotent stem cells enables analysis of glaucomatous neurodegeneration." Stem Cells doi: 10.1002/stem.2356 (2016)
- Sridhar A; Ohlemacher SK; Langer KB; Meyer JS. "Robust differentiation of mRNA-reprogrammed human induced pluripotent stem cells toard a retinal lineage." Stem Cell Translational Medicine 5:10 (2016)
- Briggs SF; Dominguez AA; Chavez SL; Reijo Pera RA. "XIST in human preimplantation embryos and iPSCs." Stem Cells 33:1771 (2015)
- 7q11.23 dosage-dependent dysregulation in human pluripotent stem cells affects transcriptional programs in disease-relevant lineages
- Josowitz R, Lu J, Falce C, D’Souza SL, Wu M, Cohen N, Dubois NC; Zhao Y; Sobie EA; Fishman GI; Gelb BD. Identification and purification of human induced pluripotent stem cell-derived atrial-like cardiomyocytes based on sarcolipin expression. PLoS ONE 9(7): e101316 (2014)
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