Amplicon Sequencing of SS3

The SS3 locus is a commonly used integration locus in E. coli engineering (Bassalo 2016). Here we validate the wildtype locus (containing the gRNA target sequence) using primers 

 

Primer Name	Sequence
SS3_forward	GGC GGT GCA CAG CAA CAC G
SS3_reverse	CCG GTG ATA CAG CTT CAA TTC CAA CC

This amplifies a short product about 250 bp long.

Interactive IGV View. After doing nanopore sequencing we can confirm the wildtype sequence with intact gRNA targeting region.

https://igv.angstrominno.com/?sessionURL=/data/AI-1047_1_SS3_locus/session.json#

Summary Table indicates that the first sample with SS3 no mutations compared to the reference.

Raw Reads (FASTQ file)

https://drive.google.com/file/d/1TZZRJAVHOC5lJUFUnrBPtQ9cGdO-Ykc0/view?usp=drive_link

Coverage Plot