Recombinant Human Turbo Dicer Enzyme Kit
Contents
1. Genlantis A division of Gene Therapy Systems Inc PRODUCT SUMMARY Cat No 1520002 50 Units Kit Components Content _ Recombinant Turbo Dicer Enzyme 1 tube x 100 ul 0 5 unit ul Dicer Reaction Buffer 1 tube x 50 ul 10 mM ATP 1 tube x 50 ul Dicer Stop Solution 1 tube x 100 ul 5X BSA 1 tube x 50 ul Nuclease free Water 1 tube x 1 ml Shipping and Storage The Recombinant Human Turbo Dicer Enzyme Kit is shipped frozen For maximum stability and long term use immediately store at 20 C upon receipt All components are stable for six months when stored properly 50 mM MgCl Solution 1 tube x 100 ul Description The Recombinant Human Turbo Dicer Enzyme Kit uses an ultra active form of human recombinant dicer enzyme that can cleave more than 95 of dsRNA template into 22 bp siRNAs within 2 hours under optimized reaction conditions Figure 1 This provides sufficient siRNAs in every single species to achieve effective gene silencing Fig 1 Efficient Digestion of Double Stranded RNA with Recombinant Human Turbo Dicer Enzyme I 3 Agarose Gel Lane Sample 1 100 bp marker 2 lug 700 bp dsRNA 3 1ug 700 bp dsRNA 15 hour digestion with 1 unit Dicer Enzyme 4 1ug 700 bp dsRNA 2 hour digestion by 1 unit Turbo Dicer Enzyme Recombinant Human Turbo Dicer Enzyme Kit Page 1 of 2 Recombinant Human Turbo Dicer Enzyme Kit NOTE The Recombinant Human Dicer Enzyme Kit is intended FOR2. For efficient generation of siRNA Stanford University The purchase of this product conveys to the Dicer siRNA Generation Kit 5 genes 50 T510001 buyer the limited non exclusive non transferable right without the transfections right to resell repackage or further sublicense under these patent TurboScript T7 20 Reactions T510003 rights to perform the siRNA production methods claimed in those es patent applications for research purposes solely in conjunction with Transcription Kit this product No other license is granted to the buyer whether RNA Purification Column 1 20 Columns T510004 expressly by implication by estoppel or otherwise In particular the RNA Purification Column 2 20 Columns 7510005 purchase of this product does not include nor carry any right or f h f th duct d lud h license to use develop or otherwise exploit this product commercially and no rights are conveyed to the buyer to use the For efficient and functional siRNA transfection product or components of the product for any other purposes GeneSilencer siRNA 200 reactions T500750 including without limitation provision of services to a third party Transfection Reagent 0 75 ml o of es e i meee or aioe i eee ai therapeutics In addition any user that purchases more than 5 GeneSilencer SIRNA gt X 200 1305750 in any calendar quarter may be outside the above research license Transfection Reagent reactions 5 x and will contact Stanford Universi
3. RESEARCH USE ONLY METHODS AND PROCEDURES Preliminary Notes 1 We recommend that you use dsRNA templates of 500 1000 bp in length The dicer enzyme might not digest well if the dsRNA is smaller than 300 bp 2 For optimal results we recommend using the GTS TurboScript T7 RNA Transcription Kit Catalog T510003 to generate double stranded RNA dsRNA dsRNAs generated with TurboScript can be used directly with the Recombinant Turbo Dicer Enzyme Kit without purification however dsRNAs purified using standard lithium chloride or equivalent procedures can give slightly better results Generation of siRNA Using Recombinant Turbo Dicer Enzyme 1 Keep the Dicer Reaction Buffer at room temperature while assembling the reaction The following amounts are for a single 10 ul reaction 3 ul x Nuclease free water x ul dsRNA 1 ug 1 ul 10 mM ATP 1 ul 5X BSA 2 ul 50 mM MgCl 1 ul 10X Dicer Reaction Buffer 2 ul Recombinant Turbo Dicer Enzyme 0 5 unit ul NOTE J Ug of dsRNA control template will yield about 0 5 ug of diced siRNA d siRNA after purification with GTS RNA Purification Columns 1 amp 2 this is sufficient for one or two transfections in 24 well plates using the GeneSilencer siRNA Transfection Reagent For more d sIRNA s scale up the reaction volume proportionally IMPORTANT Avoid using excess Recombinant Dicer Enzyme as it may decrease the amount of d siRNA that can be generated from the digestion rea
4. ction 2 Incubate for 2 hours at 37 C 3 Stop the reaction by adding 2 ul Dicer Stop Solution reVMV032206 4 Check d siRNA 22 base pairs by using either of the following two methods a 3 agarose Gel TAE b 15 native polyacrylamide gel 29 1 cast in 1X and electrophoresed in 0 5X TBE Run at 10 Watts and 4 C Visualize RNA by staining with ethidium bromide TROUBLESHOOTING GUIDE d siRNA yield is low No dsRNA or poor dsRNA Check the quality of the dsRNA produced in vitro For best results quality use the TurboScript T7 Transcription kit Cat T510003 Also check the amount of dsRNA added If purification columns were used make sure that the correct fractions were used if columns or other purification method were not used purify dsRNA before the dicer reaction Too much Recombinant Dicer Use 1 unit of the enzyme for every microgram of dsRNA Enzyme was added 10 mM ATP is old Use fresh ATP solution Low silencing effect Not enough d siRNAs used Check the amount of d siRNA produced by gel electrophoresis d siRNA quality is poor Purify the d siRNAs for best results we recommend using the RNA purification Columns 1 and 2 Cat T510004 and T510005 RELATED PRODUCTS NOTE The Recombinant Human Dicer Enzyme Kit is intended FOR RESEARCH USE ONLY Product Size Cat No Limited Label License for the Recombinant Turbo Dicer Enzyme f This product is covered by several patent applications owned by the
5. ty for a license This product is 0 75 ml sold pursuant to a license from Stanford University and Stanford University reserves all other rights under these patent rights For information on a license to the patent rights for uses other than in For efficient transfection of DNA into diverse cell lines aii ilar N conjunction with this product or to use this product for purposes GenePORTER 2 T202007 other than research please contact Stanford University at 650 723 Transfection Reagent 0 75 ml 0651 This is Stanford University reference S02 028 GenePORTER 2 150 reactions 7202015 Transfection Reagent 1 5 ml _ Please contact us for our latest catalogue of life GenePORTER 2 750 reactions 5 7202075 science research products Transfection Reagent x 1 5 ml a Genlantis For 3 minute transformation into E coli A division of Gene Therapy Systems Inc TurboCells Chemically 20 x 50 ul C300020 10190 Telesis Court ee Competent coli TETRI Chemically 20x50 ul C301020 e es ee Competent coli Tel 858 457 1919 888 428 0558 Fax 858 623 9494 Website http www genlantis com Recombinant Human Turbo Dicer Enzyme Kit Page 2 of 2 revMV032206
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