Q1) How to ship purified gDNA for species barcoding?
Ans: We recommend our customers to ship gDNA at 4oC or blue ice.
Q2) How to send samples for species barcoding?
|Purified gDNA||For all types of organisms.|
|Solution A and B||For gram-negative bacteria, fungi and plant only.|
|Solution A, B and TP||For animals only.|
|FTA Card||For gram-negative, gram-positive bacteria, animal blood only|
|0.5 x 0.5cm agar contains a bacteria/ fungi/ plant, triplicates||Samples can be shipped at room temperature|
|0.5 x 0.5cm animal/ plant tissue samples, triplicates||Please ship samples on dry ice|
Q3) What regions of the 16S rRNA gene do 1st BASE PCR in the bacterial species barcoding service?
Ans: The 16S gene is characterized by nine variable regions (V-regions) that are flanked by conserved sequences. Our 16S rRNA full length species barcoding targets on the whole 16S gene and its length is 1500bp. On the other hand, for 16s RNA partial species barcoding, we are targeting on V4-V6 hypervariable regions, resulting in a 600 bp PCR product. The latter is recommended as a survey of bacterial diversity in metagenomic studies using next generation sequencing.
Q4) The blast results did not show the expected species, is there a reason for this?
Ans: Although our species barcoding is able to identify organism at genus level, it is not always the case when it comes to species level. We recommend identifying the unknown sample with universal species barcoding primers first. After which, more specific primers can be evaluated to further categorize the organisms at species level.
Q5) Can I have the gDNA extraction method, PCR method and primer sequence of your species barcoding services?
Ans: Due to intelligence proprietary, we are not able to disclose these information to our customers.
If you would like to have the general protocols of gDNA extraction, PCR condition and the primer sequences, please order our PCR optimization services and supplying your own self-designed primer sequences for species identification
Q6) What is the alternative if the species barcoding service did not target the gene I wanted?
Ans: You can consider our PCR optimization service (MBS-2001/MBS-2002). Please provide the articles and primer sequences you are interested to work on. After which, we will synthesize the primers and perform the PCR and sequencing for you.
Q7) What is the level of accuracy for your species barcoding? Can I confirm that the species level of my sample using the BLAST result provided?
Ans: We only guarantee the accuracy of the identification at genus level. If you are looking for accuracy of identification at species level, it is recommended to have a second reference of gene or additional biochemical or morphology test to re-confirm the results.
Q8) Is there an alternative for identification of Bacillus species?
Ans: Beside 16s rRNA for bacteria species barcoding, we also provide rpoB species barcoding for Bacillus species. However, we will need to identify that the bacteria is Bacillus using 16sRNA species barcoding first before proceeding to rpoB species barcoding.
Q9) What is 18S rRNA species barcoding?
Ans: 18S rRNA species barcoding is used to differentiate between prokaryote and eukaryote ( e.g. bacteria and fungi). In addition, it is commonly used to identify microalgae species.