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Dear Researcher: You have received a series of inducible lentiviral gene silencing vectors and gene expression vectors. All of the vectors for inducible gene silencing (pInducer10’s should be gro!n in "# !ith 100 ug$%l of a%picillin. &he' all have a he) *l'+,- gene (1.- b bet!een /hoI and coRI. You can replace the he) *l'+,- gene !ith a %ir 0 based hairpin of 'our interest. All of the vectors for inducible gene expression (pInducer+0’s should be gro!n in "# !ith 20 ug$%l of a%picillin and 0 ug$%l of chlora%phenicol. I found it is easier to gro! the gene expression vectors at 0 ° 3. All of the vectors are alread' in .coli. 4pon receipt5 'ou should la' the 6lter paper on "# plate containing appropriate antibiotics and gro! overnight5 then strea for single colon' the next da'. If 'ou have an' 7uestion5 feel free to contact %e (%li8rics.b!h.harvard.edu. )incerel'5 9a%ie "i

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Dear Researcher:

You have received a series of inducible lentiviral gene silencing vectors and gene expression vectors. All of the vectors for inducible gene silencing (pInducer10s) should be grown in LB with 100 ug/ml of ampicillin. They all have a PheS Gly294 gene (1.4 kb) between XhoI and EcoRI. You can replace the PheS Gly294 gene with a mir30 based hairpin of your interest. All of the vectors for inducible gene expression (pInducer20s) should be grown in LB with 50 ug/ml of ampicillin and 30 ug/ml of chloramphenicol. I found it is easier to grow the gene expression vectors at 30C. All of the vectors are already in E.coli. Upon receipt, you should lay the filter paper on LB plate containing appropriate antibiotics and grow overnight, then streak for single colony the next day. If you have any question, feel free to contact me ([email protected]).

Sincerely,

Mamie Li