Article
Xu Zhang†,‡*
A Comprehensive Experiment for Molecular
Biology: Determination of Single Nucleotide
Polymorphism in Human REV3 Gene Using
PCR–RFLP
From the †Department of Molecular Biology and Diagnostics, School
of Medicine, Jiangsu University, Zhenjiang, Jiangsu 212013, China,
‡Jiangsu Key Laboratory of Medical Science and Laboratory Medicine,
Zhenjiang, Jiangsu, China, §Experimental Teaching Center, School of
Medicine, Jiangsu University, Zhenjiang, Jiangsu, China
Abstract
Laboratory exercise is helpful for medical students to under-
stand the basic principles of molecular biology and to learn
about the practical applications of molecular biology. We
have designed a lab course on molecular biology about the
determination of single nucleotide polymorphism (SNP) in
human REV3 gene, the product of which is a subunit of DNA
polymerase f and SNPs in this gene are associated with
altered susceptibility to cancer. This newly designed experi-
ment is composed of three parts, including genomic DNA
Keywords: Laboratory exercise; molecular biology; SNP; PCR–
RFLP
Introduction
Molecular diagnostics detect and measure the presence of
nucleic acids or proteins that are associated with health or
Volume 45, Number 4, July/August 2017, Pages 299–304
*To whom correspondence should be addressed. E-mail: xuzhang@ujs.
edu.cn or 15951285038@163.com.
**This work was supported by Jiangsu Province’s Qing Lan project,
the Research on the Reform of Higher Education Grant of Ministry of
Education (2015Y0603, JX2016-203, JX2016-Y002), the Starting
Foundation for Senior Talents of Jiangsu University (Grant No.
13JDG086), the Construction Fund for the Brand Major of Jiangsu
Province (Grant No. Yj201612), the Research on the Reform of
Education Grant of Jiangsu University (Grants No. 2013JGZD001 and
2015JGZZ008), the Research on the Reform of Higher Education Grant
of Jiangsu Province (Grants No. 2013JSJG030 and 2013JSJG280).
Disclosure—The authors have no conflict of interest to be declared.
Received 1 April 2016; Revised 31 October 2016; Accepted 29
November 2016
DOI 10.1002/bmb.21037
Published online 1 February 2017 in Wiley Online Library
(wileyonlinelibrary.com)
Biochemistry and Molecular Biology Education
Meng Shao‡
Lu Gao§
Yuanyuan Zhao§
Zixuan Sun†
Liping Zhou†
Yongmin Yan†,‡
Qixiang Shao‡
Wenrong Xu‡
Hui Qian†,‡*
extraction, gene amplification by PCR, and genotyping by
RFLP. By combining these activities, the students are not
only able to learn a series of biotechniques in molecular biol-
ogy, but also acquire the ability to link the learned knowl-
edge with practical applications. This comprehensive
experiment will help the medical students improve the con-
ceptual understanding of SNP and the technical understand-
ing of SNP detection. V C 2017 by The International Union of
Biochemistry and Molecular Biology, 45(4):299–304, 2017.
diseases, revealing the mechanisms of disease and enabling
healthcare at an individual level. Molecular diagnostics can
facilitate the practice of personalized medicine and precision
medicine. For undergraduate students in medical school, the
molecular biology course generally includes two parts: intro-
ducing the basic principles of molecular biology and the
applications of molecular biology in medical practice. Com-
pared to the other courses, molecular biology course de-
mands lab activities to help students understand the molecu-
lar mechanisms of disease and the diagnosis at molecular
level [1]. Lab activities will enable students to learn the basic
molecular biology techniques, get familiar with the general
procedure of molecular diagnostics, and gain systemic train-
ing in scientific research [2, 3]. Moreover, lab course may
include different but related activities, which will improve
the learning ability of the students [2, 4].
Genetic variation at the nucleotide level is the most
fundamental difference between two individuals, which is
commonly referred to as single nucleotide polymorphism.
Single nucleotide polymorphisms (SNPs) are abundant in
human genome and can occur approximately once every
299

several hundred base pairs. SNPs localize in both coding
and noncoding regions of the genome. SNP is emerging as
the third generation of genetic markers. SNP detection has
been used in epidemiological studies, medical examina-
tions, and drug development. SNP is one of the most impor-
tant topics for molecular biology course [2]. Although a
number of new technologies for SNP detection such as DNA
microarray and next-generation sequencing (NGS) have
been developed in the past few years, the undergraduate
students in medical school have few chance to perform
these techniques. Therefore, a lab course that allows the
students to detect SNP using traditional and affordable
techniques is required.
Laboratory techniques to analyze SNP involve the use
of DNA restriction enzymes to generate DNA molecules of
varying length depending on the presence or absence of
restriction sites. The length variations in DNA molecules
caused by restriction enzyme cutting are called restriction
fragment length polymorphism (RFLP). As a molecular
marker, RFLP is frequently used in genome mapping and
in variation analysis such as genotyping, forensics, paterni-
ty tests, and hereditary disease diagnostics. Polymerase
chain reaction (PCR) is a sensitive and efficient method for
amplifying the DNA fragment within a specific region of the
target gene. When the two techniques are combined, the
method is referred to as PCR–RFLP. To resolve and mea-
sure the length of the digested products after the restriction
enzyme reaction, a separation step by electrophoresis is
needed. Altogether, these procedures offer a rapid and reli-
able method for detecting SNP in the lab [2].
REV3 gene locates on chromosome 6q21 and contains
34 exons and 33 introns. REV3 gene encodes the catalytic
subunit of DNA polymerase f, which plays an important
role in DNA damage repair [5–7]. REV3 is reported to play
as a tumor suppressor in cancer [8]. Loss of REV3 causes
chromosomal instability in mammalian cells and enhances
spontaneous tumorigenesis [9]. Genetic variations in REV3
are associated with breast cancer risk [10]. In addition,
SNP in 30 -UTR of REV3 contributes to altered lung cancer
susceptibility through the disruption of miRNA-mediated
regulation [11]. The tagSNP rs12211763 (A/G) of REV3
gene is included in a restriction enzyme cutting site (Fig.
1). The SNP site and its flanking sequence is A/GGATCT.
There is a restriction site of Bgl II in the allele A but not in
the allele G. Therefore, The PCR product spanning this SNP
site can be distinguished by Bgl II digestion. The restriction
enzyme digestion will generate two products in individuals
with A allele, while those with G allele have one product
and the heterozygous individuals have three products.
Thus, the genotype of each individual can be easily de-
tected by PCR–RFLP.
Herein, we reported the design of a molecular biology
lab course, which includes several experiments modified
from one of our research projects. This lab course is
designed for medical students to better understand the
300
Biochemistry and
Molecular Biology Education
The principle of genotyping by PCR–RFLP in this
FIG 1 experiment. [Color figure can be viewed at
wileyonlinelibrary.com]
concept of SNP, the association of SNP with disease, and
the commonly used methods for SNP detection. Three
major parts are included in this lab course, which are
genomic DNA extraction, gene amplification by PCR, and
genotyping by RFLP. This lab course has several advan-
tages. First, this course includes a variety of experiments
that are combined in a logical and systematic manner. Sec-
ond, this experiment is performed in an exploratory way,
which will stimulate the learning enthusiasm of the stu-
dents. Third, this experiment mimics the process of scien-
tific research, which will be helpful for the students to per-
form gene-disease association study in the future [12].
Taken together, this laboratory exercise will stimulate the
medical students to understand the relationship between
SNP and disease and to learn the regular procedure for
SNP detection.
Course Information
Experimental Design
This comprehensive experiment contains three parts,
including genomic DNA extraction from human peripheral
blood samples, PCR amplification of specific region in REV3
gene, and identification of the genotype by RFLP analyses.
Materials
Peripheral blood samples were collected from healthy
donors who received medical examination in the Staff Hos-
pital of Jiangsu University. The use of human samples was
approved by the Ethics Committee of Jiangsu University.
The samples were obtained in a blinded way and no identi-
fication information was included with the samples. Geno-
mic DNA isolation kit was purchased form Qiagen (Germa-
ny). PCR reagents were provided by Takara (Japan).
Restriction endonuclease Bgl II was bought from NEB (Chi-
na). Agarose was provided by Biowest (Spain)
A Comprehensive Experiment for Molecular Biology
Methods
Isolation of Genomic DNA from Human Peripheral
Blood Samples
To save time and ensure lab safety, we used commercial
kit to isolate genomic DNA from human peripheral blood
samples. Briefly, 200 mL human peripheral blood sample
were taken into a tube that contained 20 mL protease. The
blood sample was mixed with 200 mL lysis buffer by vortex-
ing for 1 min. The mixture was incubated at 568C water-
bath for 10 min. Two hundred microliters of ethanol were
added and mixed by vortexing. All the solution was trans-
ferred to a spin column which was attached to a collection
tube and centrifuged at 10,000 rpm for 1 min. The filtrate
was then discarded. After being washed with 75% ethanol
twice, the spin column was attached to a new collection
tube and 50 mL elution buffer was added into the center of
the spin column. The spin column was recentrifuged at
10,000 rpm for 1 min to collect the eluted DNA. Finally, the
concentration of the extracted genomic DNA was measured
in a UV spectrophotometer and calculated by using the fol-
lowing formula: DNA concentration (ng/lL) 5 50 3 OD260.
The purity of the extracted genomic DNA was evaluated by
OD260/280 and OD260/230 values. The integrity of the ext-
racted genomic DNA was evaluated by 1% agarose gel
electrophoresis.
PCR Amplification of REV3 Gene
The specific primers for the amplification of REV3 gene
were designed by using Primer Premier 5.0 software. The
sequences of the primers are listed below (Forward: GCAA
CAAAGCGAGATTCCGA, and Reverse: TCTTGTCTTGTTGCA
TAGCTGT). PCR amplification of REV3 gene by using this
pair of primers generates a 312 bp product. The extracted
genomic DNA was used as the template for PCR. The com-
ponents of the PCR reaction mixture were as follows: geno-
mic DNA (200 ng), 2.5 lL 103 PCR buffer, 1 lL dNTP
(10 mM/L), 1 lL forward primer (5 lM/L), 1 lL reverse prim-
er (5 lM/L), 0.5 lL Taq DNA polymerase. Double distilled
water was added to a final volume of 25 lL. The PCR reac-
tion mixture was predenatured at 948C for 5 min, followed
by 35 cycles of PCR amplification including denaturation at
948C for 30 sec, annealing at 618C for 30 sec, extension at
728C for 30 sec. A final extension at 728C for 10 min was
performed. All the PCR products were verified by 1.5% aga-
rose gel electrophoresis. The PCR products were visualized
under a UV transilluminator and photographed.
RFLP Analyses of PCR Products
The verified PCR products were used for restriction enzyme
digestion. The PCR products that contain an AGATCT
sequence from nucleotides 66–70 will be cut by restriction
endonuclease BglII while the PCR products containing a
GGATCT sequence will not. The components of the restric-
tion enzyme digestion reaction were as follows: 10 lL PCR
product, 2 lL 103 restriction enzyme digestion buffer, 0.5 lL
Zhang et al.
BglII restriction endonuclease. Double distilled water was
added to a final volume of 20 lL. The restriction enzyme
digestion reaction mixture was incubated at 378C waterbath
overnight. All the restriction enzyme digested products were
verified by 1.5% agarose gel electrophoresis and visualized
under a UV transilluminator and photographed.
Outcome Evaluation
The written examination was used to evaluate the study.
The performance of the students on the exam questions
specifically related to SNPs was compared. The examina-
tion was designed as follows: explanation of terms, short-
answer questions, and discussion essays: 10 explanation of
terms (three point per question, that is, a maximum of 30
points in total could be obtained), 5 short-answer questions
(8 points per question were possible, i.e., the maximum
score was 40 points) and 2 discussion essays (15 point per
question, i.e., the maximum score was 30 points). The
exam questions specifically related to SNPs included one
explanation of terms, one short-answer question, and one
discussion essay (the maximum score was 26 points). Stan-
dard answers for all questions were defined by the instruc-
tor before students’ answers in the examination were
scored.
Statistical Analysis
Data are presented as means 6SD. Two-tailed t-test was
used to assess the differences of the data by SPSS software
(version 22.0).
Results
Genomic DNA Isolation and Measurement
The genomic DNA could be rapidly prepared from human
peripheral blood samples by the students in three teaching
hour. The quantity and quality of the extracted genomic DNA
were determined by using NanoDrop 1000 spectrophotometer
and agarose gel electrophoresis. The concentration of the iso-
lated genomic DNA varied from 50 to 200 ng/lL among differ-
ent groups (Table I). The total DNA yields from all the groups
met the general expectation. The purity of the prepared geno-
mic DNA was evaluated by OD260/280 and OD260/230 values as
provided by NanoDrop 1000. The genomic DNA prepared by
most groups had an OD260/280 value around 1.8 and an OD260/
230 value higher than 2.0, suggesting a good purity of the
extracted genomic DNA. The genomic DNA prepared by
groups 1 and 10 had an OD260/230 value less than 2.0, indicat-
ing the contamination of organic compounds such as remain-
ing ethanol. To examine the integrity of the prepared genomic
DNA, agarose gel electrophoresis was performed. As shown
in Fig. 2, the genomic DNA extracted by most groups could be
visualized on the gel under the UV illuminator. The prepared
genomic DNA had a size around 15 kb, indicating the high
integrity of DNA samples. The signal intensities of different
DNA samples are positively correlated to their concentrations
as provided by NanoDrop 1000.
301
 Biochemistry and
Molecular Biology Education

The quantity and quality of genomic DNA

TABLE I isolated from human peripheral blood samples
(analyzed by NanoDrop 1000)

Concentration
Group ID (ng/lL) OD260/280 OD260/230

1 46.8 1.82 1.63

2 72.5 1.85 2.27 Agarose gel electrophoresis of PCR products.
FIG 3 The PCR-amplified REV3 gene spanning tagSNP
3 101.6 1.80 2.6
rs12211763 was resolved in 1% agarose gel; 5 lL
4 113.7 1.84 2.31 of PCR product were loaded into the agarose gel
for electrophoresis. M: DL2000 DNA marker; B:
5 151.5 1.84 2.6 blank; 1–12: individual PCR product.

6 200.2 1.78 1.97

reagents. The quality of the genomic DNA used as PCR
7 231.1 1.81 2.45 template has also been verified. Problems may come from
8 201.1 1.80 2.04 the missing of some PCR components in the reaction mix-
ture or the improper mixing of genomic DNA sample with
9 115.5 1.83 2.04 the other PCR components.
10 50.1 1.82 1.74 Genotyping by RELP
11 133.6 1.81 2.45 To avoid loss of PCR products, the purification step was
skipped in this experiment. To ensure complete digestion,
12 138.8 1.84 2.46 the PCR products were incubated with restriction endonu-
clease BglII overnight. The digested products were ana-
lyzed on 1.5% agarose gel. The SNP genotype of different
PCR Amplification of REV3 Gene individuals was determined according to the profiles of
The amplified DNA fragment span the SNP rs12211763 in digested DNA in electrophoresis. The restriction endonucle-
REV3 gene. The size of the PCR products was verified by ase BglII specifically recognizes the AGATCT sequence and
agarose gel electrophoresis. All groups except group 11 got cuts the PCR products between nucleotide A and G, thus
positive results and the length of the amplified DNA frag- generating two products with the sizes of 70 and 242 bp. If
ment was consistent with our expectation (Fig. 3). The stu- the nucleotide A changes to G, the PCR products will not be
dents’ main problem in this step is pipetting errors. The cut, thus generating a product with the size of 312 bp. As
failure to get PCR product by group 11 may be due to this
reason since the PCR parameters have been optimized and
the PCR components used were from the same batch of

Agarose gel electrophoresis of genomic DNA
FIG 2 extracted from human peripheral blood samples.
Genomic DNA was isolated from 200 lL of human
peripheral blood and dissolved in a final volume of
50 lL of elution buffer. Five microliters of genomic
DNA solution were loaded into the agarose gel for
electrophoresis. M: DL15000 DNA marker; 1–12:
individual genomic DNA sample.
Agarose gel electrophoresis of restriction enzyme
FIG 4 digested products. PCR products were digested
with Bgl II overnight. As indicated, only one band
(312 bp) was detected for GG alleles, two bands
(242 and 70 bps) for AA alleles, and three bands
(312 bp, 242 bp, and 70 bps) for AG alleles. The
genotypes of sample 1, 2, 3, and 4 were AG, 5, 6,
and 7 were AA, 8 and 9 were GG.

302 A Comprehensive Experiment for Molecular Biology

 Comparison of the two groups’ examination scores. A, the total examination scores of students from two groups.
FIG 5 B, the SNP-related questions examination scores of the students from two groups. *p < 0.05.
shown in Fig. 4, only one band (312 bp) was detected for
GG alleles, two bands (242 bp and 70 bps) for AA alleles,
and three bands (312 bp, 242 bp, and 70 bps) for AG
alleles.
Outcome Evaluation Results
The comparison of the examination scores showed a signif-
icant difference between the students who participated in
this lab course (80.1 6 6.1 points) and those who did not
participate in this lab course (74.1 6 7.6 points), p 5 0.014
(Fig. 5A). Analysis of the scores in the exam questions spe-
cifically related to SNPs revealed significant differences,
p 5 0.017 (Fig. 5B); the students who participated in this
lab course (18.4 6 3.4 points) performed better than those
who did not participate in this lab course (14.5 6 3.6
points).
Discussion
In this comprehensive experiment, we combined three activi-
ties together to help medical students improve their under-
standing about SNP. We designed this comprehensive experi-
ment based on the procedure of SNP detection. In general,
10 teaching hours were used to complete this comprehensive
experiment. The students were divided into several groups
with two students in each subgroup. They used three teach-
ing hours for genomic DNA isolation and characterization,
three teaching hours for PCR amplification, and four teach-
ing hours for RFLP analysis. By performing this experiment,
the students can better understand the principles of DNA
isolation, amplification, and detection. The students also
receive trainings in multiple bio-techniques [2, 13–15].
For medical students, it’s better to let them “see” the
SNP in the lab rather than “teach” the SNP in the classroom.
In this lab course, the students isolated genomic DNA from
human peripheral blood, amplified the target gene by PCR,
Zhang et al.
and detected the SNP by RFLP. This experiment is designed
as an exploratory experiment. The students will be encour-
aged to identify the genotypes of different individuals. These
activities will facilitate the students’ learning enthusiasm.
Thus, the students’ study results will be improved through
this process of education.
We found that the students performed better on the
exam questions specifically related to SNPs after the intro-
duction of this lab course. The initial assessments of the
course indicate that the students were more satisfied with
it after the introduction of this lab course. The students
express an increased interest in learning molecular biology
and performing research. The students also express an
increased understanding of how SNPs are detected in the
laboratory. They would be better able to describe the
results of SNP analyses. This study will be conducted in a
large group of students to verify group differences. A ques-
tionnaire will be designed to collect the feedback from the
students about this lab course [16]. Moreover, the influence
of this lab course on the long-term performance of the stu-
dents will be assessed.
This lab course provides a chance for the students to
better understand the clinical application of basic science.
After they completed these experiments, the students ana-
lyzed the data and interpreted the results based on what
they had learned in the theory course. The students were
stimulated to think, to discuss, and to solve problems. For
instance, the students were asked to compare the tradition-
al approaches for SNP detection with the advanced ones.
Thus, the students had an improved understanding about
SNP by both acquiring theoretical knowledge in the class-
room and obtaining practical knowledge in the laboratory
[4, 12]. The two types of knowledge are complementary,
and facilitate a more comprehensive understanding of the
topic. The additional active learning provided by the lab
experience (a completely different learning style from
303

passive learning in classroom lectures or while reading) is
helpful to ensure that the information is learned initially
and remembered beyond the examination.
Although the students may have experience of technical
training in other courses, some common handling errors
such as improper pipetting and mixing were still present.
During this experiment, the students were given feedback
on their practical skills and given specific pointers on how
to improve by the instructor. The students also learned
how to conduct experiments in a safe and responsible man-
ner. For instance, when handling blood samples, they need
to wear gloves and had to discard the infiltrates in certain
containers.
Proper primers and conditions are important for a suc-
cessful PCR amplification. The students were introduced on
how to design PCR primers so that they can retrieve the
sequence of a target gene from the database and design
their own PCR primers. Moreover, the common problems
and the corresponding solutions for PCR amplification such
as how to reduce/avoid primer dimmers and nonspecific
PCR products were discussed in this course. The instructor
took this experiment as an example to show the students
how to optimize the PCR parameters.
The determination of genotype by RFLP is based on the
pattern of digested DNA in the electrophoresis. There are
several factors that may influence the final readout of the
results. The proper separation of the digested DNA by elec-
trophoresis is required. The instructor introduced the prin-
ciple of electrophoresis and discussed about the association
between the concentration of agarose gel and the sizes of
separated DNA. The factors that affect the migratory speed
of DNA in the gel were also discussed.
In summary, a molecular biology lab course that is
composed of several experiments is set up in this study.
This lab course not only facilitates the learning interest of
the students but also improves their study results.
References
[1] White, H. B., Benore, M. A., Sumter, T. F., Caldwell, B. D., Bell, E. (2013)
What skills should students of undergraduate biochemistry and molecu-
lar biology programs have upon graduation? Biochem. Mol. Biol. Educ.
41, 297–301.
304
Biochemistry and
Molecular Biology Education
[2] Zhang, B. Wang, Y. Xu, X. Guan, X., and Bai, Y. (2013) Using PCR–RFLP
technology to teach single nucleotide polymorphism for undergradu-
ates. Biochem. Mol. Biol. Educ. 41, 262–266.
[3] Witherow, D. S. and Carson, S. (2011) A laboratory-intensive course on
the experimental study of protein–protein interactions. Biochem. Mol.
Biol. Educ. 39, 300–308.
[4] Li, J., Shi, X., Wang, J., Song, J., Xu, L., Xu, G., Lu, L. (2015) Compre-
hensive experiment-clinical biochemistry: Determination of blood glu-
cose and triglycerides in normal and diabetic rats. Biochem. Mol. Biol.
Educ. 43, 47–51.
[5] Lin, X., Trang, J., Okuda, T., Howell, S. B. (2006) DNA polymerase zeta
accounts for the reduced cytotoxicity and enhanced mutagenicity of cis-
platin in human colon carcinoma cells that have lost DNA mismatch
repair. Clin. Cancer Res. 12, 563–568.
[6] Kotov, I. N., Siebring-van Olst, E., Knobel, P. A., van der Meulen-
Muileman, I. H., Felley-Bosco, E., van Beusechem, V. W., Smit, E. F.,
Stahel, R. A., Marti, T. M. (2014) Whole genome RNAi screens reveal a
critical role of REV3 in coping with replication stress. Mol. Oncol. 8,
1747–1759.
[7] Knobel, P. A., Kotov, I. N., Felley-Bosco, E., Stahel, R. A., Marti, T. M.
(2011) Inhibition of REV3 expression induces persistent DNA damage
and growth arrest in cancer cells. Neoplasia 13, 961–970.
[8] Brondello, J. M., Pillaire, M. J., Rodriguez, C., Gourraud, P. A., Selves,
J., Cazaux, C., Piette, J. (2008) Novel evidences for a tumor suppressor
role of Rev3, the catalytic subunit of Pol zeta. Oncogene 27, 6093–6101.
[9] Wittschieben, J. P., Patil, V., Glushets, V., Robinson, L. J., Kusewitt, D.
F., Wood, R. D. (2010) Loss of DNA polymerase zeta enhances sponta-
neous tumorigenesis. Cancer Res. 70, 2770–2778.
[10] Varadi, V., Bevier, M., Grzybowska, E., Johansson, R., Enquist, K.,
Henriksson, R., Butkiewicz, D., Pamula-Pilat, J., Tecza, K., Hemminki, K.,
Lenner, P., Fo € rsti, A. (2011) Genetic variation in genes encoding for
polymerase f subunits associates with breast cancer risk, tumour char-
acteristics and survival. Breast Cancer Res. Treat. 129, 235–245.
[11] Zhang, S., Chen, H., Zhao, X., Cao, J., Tong, J., Lu, J., Wu, W., Shen,
H., Wei, Q., Lu, D. (2013) REV3L 30 UTR 460 T>C polymorphism in
microRNA target sites contributes to lung cancer susceptibility. Onco-
gene 32, 242–250.
[12] Kloser, M. J., Brownell, S. E., Chiariello, N. R., Fukami, T. (2011) Inte-
grating teaching and research in undergraduate biology laboratory edu-
cation. PLoS Biol. 9, e1001174.
[13] DiBartolomeis, S. M. (2011) A semester-long project for teaching basic
techniques in molecular biology such as restriction fragment length
polymorphism analysis to undergraduate and graduate students. CBE
Life Sci. Educ. 10, 95–110.
[14] Schultheis, P. J. and Bowling, B. V. (2011) Analysis of a SNP linked to
lactase persistence: An exercise for teaching molecular biology techni-
ques to undergraduates. Biochem. Mol. Biol. Educ. 39, 133–140.
[15] Reinking, J. L. Waldo, J. T., Dinsmore, J. (2013) A trio of human molec-
ular genetics PCR assays. Biochem. Mol. Biol. Educ. 41, 173–179.
[16] Lian, J. and He, F. (2013) Improved performance of students instructed
in a hybrid PBL format. Biochem. Mol. Biol. Educ. 41, 5–10.
A Comprehensive Experiment for Molecular Biology