Aakriti Singh

7 Questions 14 Answers 0 Followers

Questions related from Aakriti Singh

Why are my HEK293T cells getting stressed after transfection with Calcium chloride and HBS?

We culture our cells in DMEM High glucose with 10% FBS and ABAM. However, previously we were using High Glucose w/o L-Glutamine and now the media contains L-Glutamine. Even after reviving our...

27 September 2024 2,350 2 View

I am getting double bands after in vitro transcription. What should I do to get a single band?

I am using a PCR amplified product of around 56 bp with a T7 Promoter sequence and I am getting a single band after PCR. However, after in-vitro transcription of this PCR Product, I am getting two...

28 June 2023 6,253 4 View

Recovery of my vector and insert products is very low after gel extraction. How can I increase it?

I am starting with 1.15 ug of both vector and insert for double digestion. But after gel elution I am only able to get around 3.7 ng/ul; and 10 ng/ul for vector and insert respectively in 15ul of...

29 April 2023 5,480 3 View

Has anyone used restriction enzymes from SRL?

Currently, i got SRL Restriction enzymes for performing a double digestion reaction (NdeI and XhoI). However I doubt on the functionality of the enzyme. I first performed single digestion with my...

14 April 2023 286 3 View

Multiple bands in gradient PCR. How should I resolve this issue?

Hi. I performed gradient PCR reaction using a bacterial genomic DNA as template. The annealing temperature of my forward and reverse primers are 59.4 and 61.8 degree celsius. I chose a range from...

08 March 2023 3,413 4 View

What should I add extra during the In-vitro transcription of my G rich sequence by T7 Polymerase to get RNA?

I have been trying to get RNA via in-vitro transcription of a G-rich DNA sequence after PCR amplification since 5-6 months. But i fail to observe any RNA bands under UV light after running the...

02 August 2022 965 1 View

How do I achieve proper refolding of my protein?

I am trying to purify and refold an insoluble protein from E. coli cells. I solubilized and purified the protein using 8M Urea. After that, I pooled the eluted fractions containing proteins to...

01 January 1970 8,531 10 View