Bushra Khattak

10 Questions 28 Answers 0 Followers

Questions related from Bushra Khattak

What are the reasons of getting degraded RNA?

Hi everyone, recently I am too much worry about my RNA which is always bad in sense of genomic DNA or very big problem is degradation of RNA. I try many times, I always get high concentrated RNA...

22 April 2016 2,846 3 View

Any one use QIAGEN RNase free DNase set (79254)? What about it results?

Hello, I have two questions here. 1st is it important to equalize RNA concentration for cDNA synthesis in case of gene expression? I do calculation using formula Vol1 *conc1= Vol2 *conc2 , but i...

19 April 2016 1,765 4 View

Why my RNA always degrade?

Hi, I extract RNA from different plant tissues using Trizole as well as Qiagen RNAEasy kit but when ever i extract RNA by these two methods my RNA always degrade and have genomic DNA traces....

18 April 2016 5,482 3 View

Is degraded RNA converted to cDNA by RT PCR using reverse transcriptase?

Hi, I want to ask if degraded RNA is able to synthesize cDNA by RT PCR? If yes, so when we use this cDNA and do PCR for cloning, so it affect original pcr product size etc? I want to clone one...

15 March 2016 7,373 6 View

How can we know that our vector is self ligated?

Hi, I want to know, how can we know that our vector is self ligated? I did single restriction digestion of both vector and insert with SpeI. When ever i do colony PCR i get no result, what is the...

11 January 2016 4,911 18 View

Why are some colonies larger and some are small in size?

Hello, this is colony PCR i did by using gene forward and reverse primers. My question is why some colonies are small and some are big? What kind of colonies i use for plasmid extraction and...

16 December 2015 3,366 4 View

Which vector is best for proof reading polymerase?

Hi, i want to know can we avoid use of LA taq polymerase because of not proofreading capability for ligation? Can we use another vector which is use for proofreading taq polymerase? Please suggest...

24 November 2015 9,142 4 View

Why is the DNA content very low after gel extraction?

Hi, I want to ask why after restriction digestion DNA concentration is very low when ever extract from gel? What is reason? I extract Vector and my gene of interest with SpeI using 30 ul reaction,...

04 November 2015 8,815 2 View

Any suggestion on why no colonies grow on plates?

Hi, I face problem in getting colonies on plates. I extract DNA after restriction digestion but concentration is too low than 10ng/ul so i take 20 ul of total volume of ligation and keep it 4...

02 November 2015 4,994 29 View

Why do we not use PCR product directly & how can we improve DNA extraction concentration?

Hi, I want to ask why don,t we use PCR product directly in cloning, why we prefer to extract DNA in every step? Secondly when ever i extract DNA from gel its concentration is very low, around...

30 October 2015 2,631 14 View