How many types of DNA polymerase are present in bacteria?

Now i have research about miRNA expression in ovary cancer..i used pre and post op samples from epithelial ovary cancer..i want to calculate the level expression from each others..relative expression in Pre-Op and Post-op..in final result, i want to know value from fold change in the Pre Op and Post Op..so what your suggest for the best method for me to realize that?..

Note: no control sample for this research

I used PCR to add a few nucleotides to the 3' end of a gene on a pc3.1 plasmid and produced a linear vector with 18bp overlap at 5' and 3'. Then I used DpnI to cut off the template chain, transformed the linearized vector into BL21, plated it, picked a single clone and continued to culture it in Kan-resistant LB culture medium, and then handed it over to a sequencing company. The sequencing company said that my bacteria did not grow when further expanded and could not be sequenced. So I extracted the plasmid and sent it for sequencing again, and it still showed no signal. I simply asked the sequencing company to continue testing the Kan resistance gene, and it still showed no signal. If it is because the antibiotics are degraded, then when I did the transformation, the negative control that did not transform any genes did not have any colonies, which shows that the antibiotics worked. I am very confused. If the plasmid was not successfully transformed, why can it grow in the resistance culture dish?

My primers are designed to yield 1400 bp fragment. But on the gel after PCR, I am getting sharp band at 1000 bp and faint band at 1500 bp. What are the troubleshoot options that I can apply to get my band at appropriate size?

After transformation in DH5 alpha, I got positive colonies. I have confirmed it by PCR (using an isolated plasmid of positive colonies as a template to run PCR by Takara Taq) and restriction digestion. In PCR, I got exactly the same size of band as my desired interest in the gene but did not get fall out of my gene in restriction digestion.

I have attached a gel pic of PCR and restriction digested . 20 ul of restriction digestion was put at different amount of plasmid ( 5 ul and 8 ul of 140 (C1 )and 305 ng/ul (C2 )

Molecular docking Software or online application compatible with Mac OS Sonoma 14. Thank you!

I am trying to dock a Ruthenium complex with a protein. As expected, Vina does not recognize Ru atom type and throws the following error:

"PDBQT parsing error: Atom type Ru is not a valid AutoDock type (atom types are case-sensitive).

> ATOM 3 RU UNL 1 0.177 1.341 0.016 0.00 0.00 +0.000 Ru"

I have tried to modify the parameters by adding the Ru parameters (with case-sensitive alternatives), however the error persists.

Please help me to rectify this error. I am not able to find any documentation on the same for Vina. I am performing the calculations on a LINUX OS.

Hello,

I am conducting a protein-metal docking in Autodock using an Arsenic compound and am unable to produce a proper GLG file. When I complete the autogrid I get the following error in the GLG file:

I am unsure if the problem comes from an issue with the software, parameter file, or even python? The python shell shows some errors, but I don't know if that is just because the parameterization was unsuccessful.

To solve the issue I have tried the following steps but they did not fix the error:

Thank you very much.