High Quality Microquantity cDNA Libraries are constructed using our state-of-the-art methods for cDNA synthesis and cloning to produce libraries from small amounts of starting material. Like our standard libraries, these libraries are directionally cloned into Express I and then transformed into T1 phage resistant E. coli. We guarantee at least 1×106 primary clones with ≥87% recombinant clones containing an average insert size of at least 1kb, however, we generally produce libraries of 3×106 primary clones with >95% recombinant clones containing ≥1.5kb average insert size.
We will make microquantity libraries from customer supplied tissue (<1g), cells (at least 1×107), total RNA (≥50 µg) or mRNA (≥500 ng). We provide quality controlled primary or amplified libraries.
Characteristics of a cDNA Library Constructed From a Microquantity Amount of HeLa mRNA
|mRNA (ng)||Total # of primary clones||Average insert size (kb)*||Percent recombinants||Insert size range (kb)*|
|2,000||7.3×107||2.43||100||1.7 – 5.5|
|1,000||8.0×106||2.43||100||0.9 – 5.2|
|500||6.7×105||1.75||100||0.8 – 2.9|
mRNA (ng), Total # of primary clones, Average insert size (kb)*, Percent recombinants, Insert size range (kb)**Average insert size and insert size range determined by restriction enzyme digestion of 24 clones picked at random from each library.
Quality testing: Percentage of recombinant clones and average insert size determined by gel analysis of 24 clones picked at random.
Shipment and storage conditions: Libraries will be shipped in dry ice. Storage of libraries at -80°C is recommended.