Notes+on+use+of+these+primers+12-12-12

12-12-12

-The first time these primers were used the hexL and hexR primers were used in the 1st strand synthesis. Then the InFL and InFR primers were used in the 2nd synthesis. This could result in the hexL and hexR primers priming either end of a gene and the genes could be inserted into a clone in the reverse orientation. Additionally, some clones could have two hexR primers on both sides or two hexL primers on both sides. Therefore, many of the products of these reactions would be undesired or not even capable of being cloned.

-The next time I use these primers I will only use FSHX (hexL) for the 1st strand synthesis (perhaps a poly T primer will be used as well). Then I will use IFF (hexR) and IFR (InFR) for the 2nd strand synthesis. See the diagram for this plan here: "C:\kurt\storage\CIM Research Folder\DR\2012\12-12-12\cDNA library\cDNA library random hexamer plan 12-12-12.svg"

-poly dT random hexamer primer mix will be used