How can I decide which restriction enzyme in better for restriction digestion?

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Any advice on the Xylosidase of K. marxianus?

Dear All, I am trying to find amino acid sequence of beta xylosidase of K. marxianus, but it is not present in NCBI protein database. However, sequence of A. niger's xylosidase is available. So...

07 August 2015 5,291 8 View

How can I design a primer for Xylitol Dehydrogenase and Xylose Reductase genes of Kluyveromyces marxianus?

Dear Friends, I am trying to design primer design a primer for Xylitol Dehydrogenase and Xylose Reductase gene of Kluyveromyces marxianus but when I am searching gene sequence in NCBI, it is...

31 December 2014 8,900 2 View

What Should a proper ratio between Voltage and Current while running SDS-PAGE ?

I running SDS-PAGE and I am facing a problem i.e. if I am setting voltage 100V then current is going to much higher like 400mA. If I am going through current and setting it 35 mA then voltage is...

10 November 2014 2,754 4 View

What parameters should be in the primer design when designing primer for xyloser reductase and xylitol dehydrogenase gene?

I am designing primer for xylose reductase and xylitol dehydrogenase gene. Can anyone suggest me what parameters should be their in primer designing. If anybody having protocol please provide me.

08 September 2014 4,817 14 View

What is average value of Xylitol Dehydrogenase and Xylose Reductase activity assay (U/L)?

I am performing enzyme assay of Xylitol Dehydrogenase and Xylose reductase but I don't have any reference value for that. So tell me what satisfactory value of activity form ADH and XR can be...

07 August 2014 3,681 5 View

How do you design a vector for yeast transformation?

I am beginner in vector designing. My aim is to transform yeast for xdh and xi genes. so please provide me some references and protocols.

06 July 2014 9,901 1 View

Are there any workshop on Molecular Biology?

qPCR, 2D gel electrophoresis, Gene Cloning.

05 June 2014 1,475 2 View

Why Do TDS and EC Increase with Larger Wastewater Volumes, While BOD and COD Decrease?

I have carried out MFC experiments on three different volumes, 50, 500 and 1000 mL of wastewater. Results after MFC treatment shows that TDS and EC are more in larger volumes of water i.e. TDS and...

09 August 2024 9,621 0 View

Could it be a cell culture contamination?

Hello everyone! I observed in my culture (htert-RPE1 cells) an orange- red particle at the bottom of the dish. It is visible to the naked eye as a very very small red dot. Could it be a...

09 August 2024 2,824 3 View

How to confirm the site-directed mutagenesis result without performing NGS?

I'm cloning a fragment of 3200 nts into plasmid. The cloning was successful, however, 02 amino acids were mutated. Now I want to fix these 02 aa by site-directed mutagenesis technique using...

08 August 2024 4,645 2 View

Is there any way to quantify bacterial and fungal cells in their mixed culture?

I am working in fungal fermentation of soybean meal and there is bacterial growth in them at times. I am trying to quantify fungal cell counts and bacterial cells; but I haven't been able to do at...

07 August 2024 7,535 4 View

How to normalize and take the significance of the MTT OD values with 3 replicates for the same cell-line?

Hi, I have a question about normalizing the MTT OD values for doing the statistical analysis. So, if we have 3 different plates and we call them 3 different replicates, so, first we would...

07 August 2024 8,106 4 View

Why activated CAR-Jurkat cell could not kill targets?

Previously when I co-coluture anti-CD19(FMC63) CAR-Jurkat with Raji with E:T=5:1, Jurkat can eliminate Raji in 24h. However, when I test another CAR construct, although I can dectect totally CD69...

06 August 2024 641 2 View

Are there instances where molecules with larger molecular weights exhibit greater mobility than those with smaller molecular weights?

Hi, I know that low molecular weight (MW) molecules generally tend to have higher mobility, while high molecular weight molecules tend to have lower mobility. However, in my experimental...

06 August 2024 1,495 2 View

Weak DAPI staining after immunohistochemistry - how to improve?

After immunohistochemistry of previously fixed in PFA and EtOH and then frozen 20 μm sections of zebrafish brain, DAPI staining is very weak (right) compared to the same sections stained without...

05 August 2024 9,637 2 View

Why did the authors extrapolate a phenotype that they experimentally proved in one bacterial strain across the whole genus of the organism?

I aim to be as skeptical as possible regarding whether a pair of orthologous genes results in the same phenotype in their different but related bacterial organisms under similar environmental...

05 August 2024 6,787 4 View

Why my colony PCR results of my recombinant bacterial not showing any results?

I am performing ligation of the plasmid and a target gene. The steps I have taken are: 1. Double digestion of the plasmid and target gene 2. Ligation of the plasmid with the target gene 3....

05 August 2024 2,570 3 View

NILESH KUMAR SHARMA

hey thanks aravinth, i m sure it will be helpful for me.

Michael J. Benedik

While a number of restriction enzymes have isoschizomers (i.e. other enzymes recognizing the same site) in most cases only one of those is commercially available. So use that one!

Aref Sobhkhizy

you need to choose the cheapest one and choose one of the most cost effective isoforms... I don't know whether it's important to you to save the costs or not, but if so, I would prefer to use the cheapest one .... ( of course if the results are the same )

Vikas Kumar Patel

Dear Nilesh,

pls find the attached file , it will be helpful for you for restriction primer designing and digestion with enzymes.