This is a presentation encapsulating my learnings from my research internship at Allele Life Sciences, Noida.

1. © Aryan Dugar
‘Genetic Engineering’ at Allele
Life Sciences, Noida
- Aryan Dugar
(Intern from June-July, 2019)

2. © Aryan Dugar
Reflection
I engaged in a summer research internship at the laboratory of Allele Life Sciences, located in Noida.
Under the guidance of Mr. Mrutyunjay and Ms. Loma, I was taught to conduct experiments and utilize
laboratory machines in the major fields of Molecular Biology and Genetic Engineering.
Although the summer internship program is meant for students pursuing higher education, I benefitted
from the rigor and advanced expectations of the internship. I was constantly pushed to do my best, and
my mentors - and the other interns - always treated me equally. Not only did I learn from their
experiences, but the internship ascertained my passion of becoming a biological researcher.
I am grateful for the opportunity granted to me: it has not only brought me closer to biology, but it has
enabled me to understand truly how biological questions are answered through defined procedures,
effective experimentations, and creative thinking.
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3. © Aryan Dugar
Making Solutions
I learnt how to:
(i) determine the required mass of chemicals to synthesize solutions.
𝑀𝑎𝑠𝑠 𝑟𝑒𝑞𝑢𝑖𝑟𝑒𝑑 = 𝑚𝑜𝑙𝑎𝑟𝑖𝑡𝑦 × 𝑚𝑜𝑙𝑎𝑟 𝑚𝑎𝑠𝑠 ×
𝑣𝑜𝑙𝑢𝑚𝑒 (𝑐𝑚3)
1000
(ii) determine the required volume of a particular solution to synthesize a
new solution.
𝑀𝑜𝑙𝑒𝑠 𝑟𝑒𝑞𝑢𝑖𝑟𝑒𝑑 = 𝐶𝑜𝑛𝑐𝑒𝑛𝑡𝑟𝑎𝑡𝑖𝑜𝑛 1 × 𝑉𝑜𝑙𝑢𝑚𝑒 1
= 𝐶𝑜𝑛𝑐𝑒𝑛𝑡𝑟𝑎𝑡𝑖𝑜𝑛 2 × 𝑉𝑜𝑙𝑢𝑚𝑒 2
𝑴 = 𝒄 𝟏 𝒗 𝟏 = 𝒄 𝟐 𝒗 𝟐
(iii) synthesize a solution of CTAB buffer
CTAB Buffer Solution Synthesis
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4. © Aryan Dugar
Plant DNA and RNA Isolation
Procedure for Plant DNA isolation (1)
I learnt how to:
(i) utilize laboratory equipment, such as mortar
and pestle, water bath, pipette, vortex
machine, and centrifuge.
(ii) accurately measure and transfer solutions.
(iii) distinguish between supernatant and pellet.
Procedure for Plant DNA
isolation (2)
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5. © Aryan Dugar
Notes on Using Centrifuge
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6. © Aryan Dugar
Quantitative Analysis of DNA
I learnt how to:
(i) utilize the spectrometer,
(i) assess the DNA
concentration, percentage
yield, and optical density,
by making absorption
measurements at
wavelengths of 260nm and
280nm.
Readings obtained for samples 𝝀𝟏 𝒂𝒏𝒅 𝝀𝟐.
Equations to determine various properties.
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7. © Aryan Dugar
Qualitative Analysis of DNA (Agarose Gel Electrophoresis)
Failed attempt
I learnt how to:
(i) Create the agarose gel and the wells, and load DNA +
dye samples into the wells.
(ii) Qualitatively determine the presence of DNA in
particular samples through UV transilluminator.
(iii) Extract the DNA fragment from the gel (DNA
Elution).
Successful attempt
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8. © Aryan Dugar
cDNA synthesis and PCR (Polymerase Chain Reaction)
I learnt how to:
(i) use the pipette correctly, and ensure
accurate volumes of the solution
(ii) Independently operate the PCR
machine, and understand its
functioning.
(iii) handle laboratory chemicals
appropriately.
http://www.allelelifesciences.com/research-facilities.html
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9. © Aryan Dugar
Bioinformatics – Using ‘Gel Analyser’
I learnt how to:
(i) operate the ‘Gel Analyser’ software in order to interpret
data from gel electrophoresis.
(ii) generate and interpret graphs on the software.
http://www.gelanalyzer.com/
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10. © Aryan Dugar
Bioinformatics – PerlPrimer and Snapgene
I learnt:
(i) the relevance of bioinformatics software – PerlPrimer and Snapgene.
(ii) to utilize these software for simulating experiments, such as virtual PCR and
insertion of a gene of interest in a plasmid (puc19) using restriction enzymes.
(iii) the factors involved in selection of primers, such as:
- Length
- GC%
- GC content
- Self-annealing
- Melting temperature
(iv) to navigate NCBI’s Database and obtain FASTA sequences of genes, proteins and
mRNA.
http://perlprimer.sourceforge.net/screenshots.html
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11. © Aryan Dugar
Recombinant DNA
I learnt:
(i) the procedure behind synthesizing recombinant DNA
Ligation of adaptor to gene of interest (GOI)
Plasmid isolation from bacterial culture
Restriction digestion of plasmid and GOI
Ligation of GOI and Plasmid
(ii) to utilize the pipette to make accurate measurements
(iii) to carefully handle solutions and the solutions that I create, such
that mixing up is avoided.
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12. © Aryan Dugar
Synthesizing transformed bacteria
I learnt:
(i) to use the autoclave to sterilize used culture plates, LB solution etc.
(ii) to use the pH meter in order to obtain solutions of accurate pH.
(ii) to synthesize LB growth medium.
(iii) to create culture plates, and inoculate them with bacterial cells.
(iv) to produce competent cells with altered cell walls for transformation.
(v) to screen for transformed bacterial cells via:
- Blue-white screening
- Antibiotic sensitivity
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13. © Aryan Dugar
quantitative PCR (qPCR)
I learnt:
(i) the principles on which qPCR works
- Use of two DNA samples (for relative
quantification of under/overexpression of genes)
- Use of primers for both the target gene (here, Gpx
in plants) and a housekeeping gene (here, beta-actin)
(ii) how to interpret data obtained from qPCR
- Conducting calculations using Ct (threshold cycles)
values
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14. © Aryan Dugar
Protein Isolation and Estimation
I learnt:
(i) to utilize the spectrometer
and centrifuge to isolate
and estimate protein
content of lentils (using Folin-
Lowry method).
(ii) to process spectrometric
data via a calibration curve,
to determine the
protein content of the sample.
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15. © Aryan Dugar
THE END