Why I got non specific band after PCR ?

I did PCR reaction without using any PCR kit . The result is not good enough. I got heavy low non specific band and probably primer dimer (picture attached). First, I did with annealing...

09 October 2019 1,794 4 View

Why I failed to get good band after plasmid isolation?

I tried to isolate plasmid DNA from Agrobacterium tumefaciens using both manually and using kit and performed the electrophoresis on 0.5%, 0.8% and 1.0% agarose gel. The ladder showed the...

08 September 2019 2,097 2 View

How can I solve primer dimer problem without changing primer?

For primer dimer there are many reasons. One is primer designing. Is there any way to solve the problem with the same primer that have been designed?

05 June 2018 3,619 2 View

How this can be interpreted?

After PCR followed by gel run every time I got this picture. Can anybody tell me why this happened and what will be the solution? In these two pictures, one gel had been run after putting 1kb...

04 May 2018 5,260 0 View

What is the difference between PCR and gradient PCR?

Any body can explain about the PCR and gradient PCR technology using an example?

11 December 2017 4,964 2 View

Any conferences in Biotechnology before Feb 2018?

09 October 2017 2,468 2 View

What does it mean by primer length 20mer?

I designed some primer for my experiment and I do not know the actual length of the primers which I must need to know for getting the result. I got a manual with the primers supplied by the...

09 October 2017 1,948 4 View

Preparation of water extract molecule

While I am preparing WEB (Water extract molecule from Streptococcus pneumoniae) after 6 hours shaking it supposed to centrifuge in 10000rpm. But my centrifuge machine was shown error that is why I...

01 January 1970 2,828 2 View

Waters 2707 HPLC autosampler problem?

I am using a 2707 waters HPLC device. When I try to inject a sample, it says missing plate or rack. I changed the needle and calibrated its position but I still get the same problem. I even get...

02 March 2021 1,408 1 View

Should we change media after1st 6hrs after plating HUVEC bought from INVITROGEN?it's not recomended by invitrogen.Will DMSO present in stock matter?

We have HUVEC bought from Invitrogen. Cat no -C-003-5c. while first plating the cells we have bought the protocol suggests not to centrifuge the cell with media to remove the DMSO. Should we...

02 March 2021 3,713 2 View

How to normalise data on OPM package from R Studio ?

Good afternoon, I recently used OmniLog from BIOLOG for my experimentations : I tested the metabolism of different strains on 2 types of plates. I have 16 strains of 3 different groups...

02 March 2021 3,584 1 View

Benefits of the 24 welled plate?

We have always used the 96 welled plate, the 48 welled plate but apparently there is much evidence on the 24 welled plate and above all its benefits.It states it is the same you would use on ELISA...

28 February 2021 1,149 1 View

Can bacterial suspension be pipetted directly onto petri dish and agar poured onto it?

Hi, recently I encountered a protocol for plating which was not the usual method I have been practicing (pour LB agar onto plate, allow it to solidify, pipette bacterial suspension onto plate,...

28 February 2021 7,292 3 View

What is the best surface to culture cells isolated from a skin biopsy?

Hello, recently I am having some difficulties with maintaining the cell culture of primary cells isolated from a skin biopsy (normal and malignant). The coating of culture plates with Matrigel (a...

26 February 2021 6,151 2 View

Difficulty in cloning of 1kb gene into vector?

I am facing difficulties in cloning a 1kb gene into a vector (pJIT163). I have my gene on interest (GOI) in pUC57 and want to clone in pJIT163 using SalI and BamHI restriction sites. I am getting...

25 February 2021 3,221 7 View

Can cells be reused after using WST 8/ CCK 8 from Sigma Aldrich?

Hi all, Can cells be reused after I have seed them onto 96 well plates and test them with cell counting kit at OD450 (CCK8) at 12 hours, wash them with PBS x3 and add new media and let them...

24 February 2021 5,072 1 View

Standard curve shifting over time. How do I stabilise the ELISA?

24 February 2021 1,365 3 View

Is it harmful to breed for Cre Homozygosity initially?

Hello, I am currently using a Cas9 knock in system that will be expressed in the liver. To do so STOP codon fl/fl Cas9 KI mice were bred with Albumin-Cre mice to generate heterozgous pups, that...

24 February 2021 5,759 3 View