New answers tagged dna-sequencing
This depends completely on the quality of the DNA. Since each chromosome is essentially a very long strand of DNA, breakages and missing sections are very common in extinct species due to degradation over time. If a full DNA read is absent, no determination of chromosomal number can be performed. Assuming a full read (covering all breakages) of the DNA ...
You can use the PCR purification kit for cleaning up the DNA. You can use the primers that you used for RTPCR for sequencing. The universal primers would work only if the sequence complementary to it is there in the 3' end of your amplicon.
See this document on how Illumina sequencing works. When you are doing a single end sequencing, the reverse strands produced after the bridge amplification + denaturation (step-7) are specifically removed (clipped at the junction of primer and the "insert" region). Furthermore, the free 3' ends are blocked (by chemical modification) to ensure that there is ...
Presumably they removed one set of fragments from their figures for clarity of demonstration. In reality, both fragments remain immobilized on the chip. The sequencing primer only anneals to one adapter which allows selective extension of one of the sets in each run. For fragments of sufficient length, this provides an easy mechanism for creating paired end ...
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