摘要:1. Dissect embryos and place each yolk sac in a microfuge tube (can be frozen at -20°C prior to extraction). Original method cited for mouse toes, or 2 mm tail, Reagents: A) 25 mM NaOH / 0.2 mM ED[阅读全文:]
摘要:This is how we test mice and rats for the presence of the transgene by PCR. It is provided for those investigators who are unfamiliar with DNA genotyping and who need to establish a genotyping method.[阅读全文:]
摘要:1. Obtain tail biopsies from 2 to 3 week old mice: Hold mouse firmly at base of tail with one hand, with the other cut off 0.5 to 1.0 cm of the tail tip with a scalpel or single edge razor blade. 2. A[阅读全文:]
摘要:1.Cut 1/2" - 3/4" of mouse tail. (Check with your institution's Animal Studies Committee for their recommendations as to how this procedure should be implemented). 2.Add tail fragment (or tissue of th[阅读全文:]
摘要:1. Obtain the last 2mm of tail and place directly into 200ul 1X PCR Buffer with Nonionic Detergents (PBND) in a 1.5ml microfuge tube. (Tails can be stored at frozen in PBS or PBND until use.) 2. Add 1[阅读全文:]
摘要:PCR screens must be designed to detect transgene DNA at the single copy level. Southern Blots analysis of transgenic mice need copy standards to estimate copy number. Copy standards are prepared by mi[阅读全文:]
摘要:This is how we test mice and rats for the presence of the transgene by PCR. It is provided for those investigators who are unfamiliar with DNA genotyping and who need to establish a genotyping method.[阅读全文:]
摘要:These procedures were originally devised in Richard Palmiter's lab for use with tail dots (DNA spotted onto a nitrocellulose filter and probed for a transgene; quicker than doing southerns), and subse[阅读全文:]
摘要:AP Buffer:100 mM Tris-HCl, pH 9.5 100 mM NaCl 10 mM MgCl2 PTM:PBS 0.1% Tween-20 2 mM MgCl2Since this is generally done in conjunction with lacZ staining, embryo/tissue is usually fixed as per lacZ sta[阅读全文:]
摘要:NOTE: THIS IS FINE FOR SOUTHERNS, BUT NOT FOR SCREENING BY PCR. (from Ruixia, 7/99, from protocol by Stef Oehen 7/94). 1. Put 1 cm tail in 1.5 ml microcentrifuge tube. 2. Add 500µl Tail Buffer a[阅读全文:]