|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||31865||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
This material is available to academics and nonprofits only.
- Backbone size w/o insert (bp) 3956
Growth in Bacteria
Copy numberHigh Copy
Gene/Insert name24xMS2 Stem Loops - Stable
Insert Size (bp)1300
- Cloning method TOPO Cloning
- 5′ sequencing primer M13F
- 3′ sequencing primer M13R (Common Sequencing Primers)
The Singer Lab no longer recommends this plasmid, as it has a higher rate of bacterial recombination.
For mammalian cells they recommend the following plasmids:
pUbC-FLAG-24xSuntagV4-oxEBFP-AID-baUTR1-24xMS2V5-Wpre (Plasmid #84561)
HaloTag-bActinCDS-bActinUTR-MS2V5 (Plasmid #102718)
For yeast cells they recommend the following plasmids:
pET246-pUC57 12xMS2V6 (Plasmid #104390)
pET259-pUC57 24xMS2V6 (Plasmid #104391)
pET251-pUC 12xMS2V6 Loxp KANr Loxp (Plasmid #104392)
pET264-pUC 24xMS2V6 Loxp KANr Loxp (Plasmid #104393)
This version of the 24xMS2 stem loops has been engineered to allow stable propagation in bacteria. Loss of repeats is still possible, but occurs significantly less often than with previous version. This plasmid is considered the replacement for pSL-MS2-24X (Addgene plasmid# 27120).
The 24xMS2 stem loop cassettes in this plasmid were generated from two non-identical stem-loops to reduce unnecessary redundancy and the chance of recombination in bacteria. The sequence for a single cassette (with the two non-identical stem-loops in uppercase) is: tacggtacttattgccaagaaaGCACGAGCATCAGCCGTGC ctccaggtcgaatcttcaaaCGACGACGATCACGCGTCGctccagtattccagggttcatc
For your Materials & Methods section:pCR4-24XMS2SL-stable was a gift from Robert Singer (Addgene plasmid # 31865 ; http://n2t.net/addgene:31865 ; RRID:Addgene_31865)
For your References section:Localization of ASH1 mRNA particles in living yeast. Bertrand E, Chartrand P, Schaefer M, Shenoy SM, Singer RH, Long RM. Mol Cell. 1998 Oct . 2(4):437-45. 10.1016/S1097-2765(00)80143-4 PubMed 9809065