Polymerase Chain Reaction

In vitro method for producing large amounts of specific DNA or RNA fragments of defined length and sequence from small amounts of short... | Contact experts in Polymerase Chain Reaction to get answers

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Questions related to Polymerase Chain Reaction

In anyone aware of a mouse genotyping optimized protocol?

Hi, currently in our lab theres a lot of transgenic mice model generation, wich means a lot of breeding and lot of genotyping. We are using tails, we lyse them ON and run a PCR and a gel. The...

22 September 2023 6,671 2 View

Does repeated use of agarose gels affect the pre-staining gelred in the gel?

I only used 2 wells of 12-well gel for my first electrophoresis. Can I reuse the remaining wells at the end of this electrophoresis? Could the gelred dye in the gel have been affected by the...

21 September 2023 9,772 1 View

EMSA：Will the protein block the Fluorescence signal of the probe?

I'm running EMSA and add the FAM Fluorescent Groups to 5' ends of my probe. However, the cis-element of the protein in the probe is close to 5' ends(only 7bp).Will the protein block the...

18 September 2023 9,541 2 View

Why is DNA extracted from the EasyPrep liquid (PAP test) really little?

When I isolate DNA from cervical cells in EasyPrep liquid (cervical swabs), the DNA amount is very little in spite of a large quantity of cells (a big pellet at the bottom of a 1.5ml tube). The...

18 September 2023 1,425 4 View

How much PCR product do I have to use for a Cas9 digestion?

I needed to do a Cas9 digestion of the amplicon, but I did not achieve it. Some protocols refer to the use of 1-2 uL of PCR product directly with optional purification, but I do not understand why...

17 September 2023 3,511 0 View

Any ideas for research in the hematology field for startup students?

I'm a new student and I need ideas that help me in the future in Hematology or Blood Transfusion but I need it in a lab filed so I can collect data easily.

15 September 2023 9,496 7 View

I need a protocol to reduce any oxidized (Fe3+) hemoglobin in my sample to (Fe2+)?

Hi everyone, I’m using hemoglobin in the lab and I need it to be in the reduced state (so it is able to bind with CO2). The hemoglobin I have is a lyophilized powder from Sigma Aldrich (H7379)....

14 September 2023 9,101 4 View

Anybody use SFM4000/s for stop flow measurment?

I was trying to measure kinetics with our iron complex with different oxidant and substrates using double mixing mode. I don't have any previous experience with this instrument. I am always...

14 September 2023 677 2 View

How to desing “splinkerette” adapters for TraDIS (transposone) libraries?

I'm trying to optimize TraDIS for our model bacterium and I am having some technical problems with the DNA preparation for the sequencing step. I'm struggling with efficient PCR enrichment of...

13 September 2023 5,808 0 View

The isolation of genomic DNA from fermented milk samples has resulted in smears, what could be the cause?

Hi all, there have been some challenges with regards to genomic DNA extraction directly from fermented milk samples. Any input with knowledge on troubleshooting would be appreciated. I have been...

08 September 2023 380 2 View

What are the key parameters routinely measured in a hematology lab's complete blood count (CBC) test, and what clinical insights can they provide?

list the usual test done in a CBC lab test and state its purpose.

07 September 2023 7,965 3 View

Who uses Tetra-Arms PCR method ? for SNPs detecting ?

Who uses Tetra-Arms PCR method ? Who can help us in using the method specific in using primers(outer & Inner Primers) ??

04 September 2023 9,971 5 View

Trouble in Arabidopsis growth?

My plants were growing normal in starting days but after some days they got bleached. I change the sterilization protocol and media composition also but not getting appropriate results. Does...

04 September 2023 3,084 1 View

Can CRISPR eliminate food insecurity from all around the world.?

Can CRISPR play a major role to overcome the world food insecurity issues?

02 September 2023 3,564 1 View

Has anyone used the NEB Gibson Assembly kit before and have tips?

My lab has the NEB Gibson Assembly kit (unfortunately not the Hifi kit) and I have been struggling for months to assemble various different constructs. I'm wondering if anyone has used this kit...

31 August 2023 4,931 1 View

What should I do to compare many samples treated with decreasing concentrations of a compound with a single control group?

It’s OK if the answer will be Dunnett’s test. Multiple comparisons lead to alpha inflation (increase in type I error rate) so correction is required. Many-to-one comparisons are comparisons of...

30 August 2023 1,988 2 View

Although the ladder transfer is okay, I can not see my proteins on the membrane, what is the reason ?

Hello, I have been doing western-blot experiments for 8 months or something (Invitrogen B1000 Mini Blot Module). I used to get good results until 2 weeks ago. Then something happened, I could not...

30 August 2023 6,981 3 View

I am trying to review and investigate studies in using designer Cellulosomes in compost degradation, can you recommend studies on this?

I am trying to follow some established protocols in improving the lignocellulosic compost degradation using designer cellulosomes. However I struggle to find any papers directly related to this....

24 August 2023 5,324 2 View

How to check microRNA primers using NCBI ?

I will perform gene expression for mir27a and mir155 so I need to check my primers by any suitable software thank you

22 August 2023 9,900 4 View

How to improve knockout efficiency in CRISPR screen system?

I have try to establish a CRISPR screen in primary T cells，sgRNA transduce with lentivirus and Cas9 protein delivered by electroporation (SLICE). But in the pilot experiment, we found that the...

22 August 2023 5,181 2 View

What possibly a problem if i got a very high RMSD value more than 1 nm? (>10A) meanwhile it supposed to be lower than 0.2 nm or 0.3 nm?

I have run a MD simulation of complex protein SIB1 (PDB ID: 7EAU) with several small molecules (ligand) using GROMACS and all of them show huge value of RMSD. Here i attached a google drive for...

09 August 2023 7,349 10 View

Can we prepare hydrazine hydrate buffer solution? Please provide reference.?

Can we prepare hydrazine hydrate buffer solution? Please provide reference.

03 August 2023 995 3 View

What could be the reason for observing a band in NTC in RTPCR?

I have been getting Ct values almost similar to a test sample in NTC wells. The melt curve also overlaps with the test samples. While running a gel, a band with a same size of product to the...

25 July 2023 3,904 0 View

I made a lytic reagent for 3-part cell counter. How can I reduce the hemoglobin percentage?

All parameter is ok but hemoglobin is 4% higher than control. the formula is SDS + Triton x100+ phosphate buffer.

12 July 2023 8,240 2 View

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