Greeting to all,
I have tried to perform a standard curve using the Zymobiomics microbial community standard. Unfortunately both of the set of primers that I have used did not worked.
Set 1. 5=-GAT TAG ATA CCC TGG TAG TCC AC-3=
5=-TAC CTT GTT ACG ACT T-3=
Set 2
5- ACT CCT ACG GGA GGC AGC AG 3
5-ATT ACC GCG GCT GCT GG -3.
It will be extremely helpful for my experiments =, If somebody has used this microbial standard to let me know the set of primers used.
Thank you in advance
Hello,
Did you use the DNA standard or the prep standard? On the prep standard a you have to do a DNA extraction before doing the PCR. In case of using the prep standard, maybe something went wrong in the DNA extraction?
And have you successfully used these primers before? Otherwise some optimization might be needed to get the PCR working.
Cheers
Thank you so much for your reply,
I have used only The DNA standard from Zymo biomics and purchased DNAs for Treponema Denticola and Pseudoramibacter Alant. I have been able to obtain the standard curves for the bacteria but I have issues with the monk Zymo population. Have you ever used it and if yes could you please let me know the 16S universal primers set that you used with the zymo biomics monk population.
Please.
We have use the ZymoBIOMICS Microbial Community DNA Standard (D6305) in the past as a Mock community for NGS (16S).
The primers we use are:
Set 1: longer amplicon
341F 5'-CCTACGGGNGGCWGCAG-3'
805R 5'-GACTACHVGGGTATCTAATCC-3'
Set 2: shorter amplicon
515F 5'-GTGCCAGCMGCCGCGGTAA-3'
806R 5'-GGACTACHVGGGTWTCTAAT-3'
Annealing temperature of both sets at 60°C.
Hope this helps.
Could you please type again the primers set because there are some letters that do not correspond to bases
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