Notes on use of these primers 12-12-12

2015-01-13

azim58 - 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