转基因实验

Culture should be 2 days old and colonies should be of medium to large size. This is important as cells need to be in log growth phase. Two small (25cm2) flasks are required one is harvested in morni ...

Pre-sterilize two pairs of fine forceps one pair iris scissors and one blunt-ended 23 gauge needle. Take 3.5 day pregnant mouse kill and dissect out entire uterus using sterile technique. Place uteru ...

Before you start have ready the following: - Sterilize 2 pairs of curved forceps 1 pair of iris scissors and 2 pairs of fine watchmaker forceps. - 6ml syringe filled with DMEM with HEPES with an 18 ...

Blastocyst transfer is usually performed 24 hours after aggregation when the morulae have become expanded blastocysts and on the same day as injection. A little time is given between injection and tra ...

Solutions

This method works well for 17.5 day mouse embryos. All stains should be made up fresh. Fix embryos in 90% ethanol for at least 1 week longer if possible. Prior to staining remove skin and viscera pa ...

The aggregation method for generating chimaeras as opposed to the microinjection technique is useful as it does not require expensive microinjection apparatus or sophisticated manipulative skills and ...

Blastocyst transfer is best performed after allowing injected embryos a little recovery time in culture. This allows better evaluation of the cells' survival - the embryos should recavitate if they ha ...

完整的实验方法请点击下面的链接下载： ...

DNA isolation for Southern analysis 完整的实验方法请点击下面的链接下载： ...

Mouse embryos staining including: in vitro transcription of riboprobe embryo fixation and permeabilization hybridization and post-hybridization washes immunodetection and development of signal. 完整的实验 ...

FGFR-AP-FGF Binding Assay PCR Assays for Knockout Mice RNase Protection Assay:The bound receptor was quantified by measuring the initial rate of substrate hydrolysis at 405 nm and same amount of prote ...

The purity of the transgene DNA used for microinjection is critical for the successful production of transgenic founder mice. DNA impurities in the form of bacterial endotoxins or organic contaminants ...

1.Restrict DNA and gel purify Cut 100ug DNA removing as much plasmid sequence as possible from the insert. Run on an agarose gel. Cut out band and electroelute in 0.5X TBE. Add an equal volume of ...

The success of DNA purification can have profound consequences on the success or failure of any injection effort. This summary of techniques from several different sources was compiled by Brad Preston ...

We have found the UltraClean™ GelSpin™ Kit (available from Mo Bio Laboratories Catalog number 12400-250) is a simple and fast way to obtain microinjection quality DNA. This is not to exclu ...

1. Purify plasmid from bacteria. We recommend the Qiagen EndoFree Plasmid Maxi kit for the purification of the targeting vector plasmid from bacteria. Please follow the directions in the kit. Electro ...

Buffer composition is 10 mM Tris-HCl pH 7.5 0.1 mM EDTA 30 microM spermine 70 microM spermidine 100 mM NaCl. This buffer is more likely to produce transgenic mice with intact unfragmented DNA molecule ...

完整的实验方法请点击下面的链接下载：

完整的实验方法请点击下面的链接下载： ...