Help! Docker version run error when design primer using "-embedded"!

[HFyange]

I am using docker version on a Mac. I have tried the multiplex primer design by your readme text. It's amazing and very OK except when design embedded primer just adding a command "-embedded".

Please help checking the problem as described bellow:

1. The regions name is cs_glf_r.csv. Directory is Primers/cs_glf_r.csv. The region content is: chr7 55259515 55259515 L858R 1 B NotW chr7 55241707 55241707 G719S 1 B NotW chr7 55249071 55249071 T790M 1 B NotW

2. Run the following commands: python3 NGS_primerplex.py -regions Primers/cs_glf_r.csv -ref hg19/ucsc.hg19.fasta -embedded -minampllen 90 -optampllen 120 -maxampllen 130 -blast -snps -nucs 10 -dbsnp hg19/common_all_20180423_hg19.vcf.gz -primernum1 10 -minprimermelt 59 -maxprimermelt 61 -optprimermelt 60

3. There is a KeyError in the running result: Creating primer3 parameters for external primers design, combination variant 1... Constructing external primers... 11.11%Traceback (most recent call last): File "NGS_primerplex.py", line 3707, in leftGC,rightGC,outputInternalPrimers[curRegionName][7]-(start+primersCoords[0][1])+1, KeyError: 'EGFRG719S'

@aakechin Thanks to the author.

[HFyange]

Another qusition: How to design embedded primer (nest primer) for anchored PCR. Please show the command example. Thank you!

[aakechin]

@HFyange Thank you for the question. It was due to old version in the Docker. Try to use the new version uploaded: aakechin/ngs-primerplex:full_1.3.3. I tried it with your parameters, and all steps completed normally.