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While the fluorescence analysis of the ligation of these proteins demonstrates maintenance of function it is therefore of interest to demonstrate that more fragile proteins can be ligated to solid supports while maintaining function.
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The resulting DNA fragments were ligated to adaptors for the SOLiD sequencer (Life Technologies, Carlsbad, CA) for 20 minutes at RT using the Quick Ligase Kit (NEB), followed by purification using the MinElute Reaction Cleanup Kit.
Step 2: 4.5 µg of the DNA was ligated to 1.5 µg of the 3'loop.
The restriction fragments were ligated to adapters and amplified by PCR.
Purified DNA fragments were ligated to SmaI digested and dephosphorylated pCre2.
The short fragments were ligated to Illumina sequencing adaptors.
Adenylated DNA fragments were ligated to Illumina TruSeq multiplexed adapters.
Subsequently, the short fragments were ligated to sequencing adapters.
Next, this ligated high molecular weight DNA was digested by RsaI, and the digested DNA was ligated to "Second linkers".
After adenylation of 3' ends of DNA fragments, Illumina PE adapter oligonucleotides were ligated to DNA fragments.
Subsequently, cDNA fragments were ligated to adapters after an end repair process.
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