How big is the plasmid size? Try to visit some web sites to read how genomic DNA used for YAC is isolated.DNA run on agarose gel should be quite clean with minimum junk before DNA can be separated and viewed well, whether it is conventional gel or PFGE.
The PEI polymers will have DNA condensed to form nanoparticles which are used for transfection. There are two ways to assemble nanopartices, 1) very small volumes of PEI, usually 5-10 ul 10% PEI, is added into large volumes of plasmid DNA, usually 100-200 ug plasmid DNA in a volume of 2-5 ml; 2) large volumes of plasmid DNA, that is 2-5 ml of plasm
You may need to order oligomer template and PCR primers for producing microRNA, or producing shRNA. Then you may need to make mammalian expressing vector producing the shRNA, or generating lentiviral vector producing the shRNA. After testing in cells, you can prepare concentrated lentiviral particles for direct tissue injection. If the mammalian ex
The issue is that there is no sustained levels of the transgene product although the adenovirus work fine in the testing cells. In your experiments there is no problem with protein extracts, the antibody against the trasnsgene product shows great for liver sample but protein extracts made from adenovirus infected cells in Western blot. You ma
Elevated transgene mRNA could cause elevated levels of gene procusts, or may cause transgene product level inhibited too due to too much mRNA induced cell toxic effect.In your Western blot, you should probe beta-actin or other housekeeping gene products to determine if the protein extracts relaible or not before examining the transgene product sinc
最近做了高通量测序， in NCBI gene bank data base try to tun BLAST program using the DNA sequences or cDNA sequences against genome data base. The other approach, since 结果中只给了对应的基因组位置, you can retrieve this specific genomic DNA and try to access transcriptome of htis specific genomic DNA.
酶切后还进行了验证，how do you know that the 1.3 kb fragments are cut with Mlu I + Bgl II?但是连接后就成了2.4kb左右: is this 2.4 kb fragment cut off from the vector after ligation, transformation and plasmid DNA isolation?What is the vector?